Selecting genes to patentMark Adamsa private company's approach to patenting genes

It was never our intention to patent all the genes, all the genes aren't patentable, and the PatentOffice has made that very clear, in fact, that they have stringent rules for what's patentable andwhat's not, and that neither the entire genome nor the entire set of genes would be patentable. Sowe've taken the same very selective approach to doing that, that is common in the biotechnology,pharmaceutical industries, as well as in academic universities and the NIH [National Institutes ofHealth]. After all, the NIH holds more gene patents than any other organization. It's sort of a bundle,the gene sequence and the protein and antibodies made from that protein are all typically part of apatent application, depending on what the commercial plan rationale is for it.

Billions of basesMark Adamsthere are 2.9 billion letters in the human genome

In the human genome there are 2.9 billion As, Cs, Gs, and Ts. If you wanted to readout the genomesequence and could do that at ten bases a second, ACGT, ACGT, ACGT, it would take eleven years toread the genome sequence. It's a tremendous amount of information and if you print it up on a bigwall poster, a tiny fraction of it, it's all As, Cs, Gs, and Ts.

Mutations & cancerBruce Amescancer is caused by an accumulation of mutations

So I think cancer is a disease of DNA. Each cell is programmed when to grow and when not to grow,everything's… there are all these circuits, and if you mutate some gene and upset one of thesecircuits, then the cell grows when it's not supposed to, and you end up getting a tumor cell. Andpeople have worked out there's several steps along the way, it isn't just one hit. Cancer’s severaldifferent genes you have to mutate in sequence. That’s why cancer goes up with the fourth or fifthpower of age. There’s very little cancer in young people and that's usually because you inherit amutant gene from your mother or your father, but other than that it goes just up with age because it'sseveral hits.

Weeding out diseaseBruce Amespredictions for gene testing

If you have a choice of children and some of them are smarter or some of them aren't going to die atthirty or aren't going to die in infancy, of course people will choose the embryo, the fertilized embryoto use that has better prospects. It'll start with weeding out horrible genetic diseases that people

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DNAi DVD Transcripts

know are in their family, and then there'll be, already, my daughter lives in London and she'sexpecting a baby, you have an amniocentesis or something where they test for a few, Down syndromeand a few other genetic diseases. I mean that's going to become more and more, easier and easierand it'll slowly creep in and we'll have to decide what, and people I'm sure will discuss this at greatlength, what's the right thing to do and what isn't and all of that.

Lab safetyEmmett Barkleydemonstrating the P4 lab containment suit he developed for working with high risk substances

The most extreme form of containment was called P4. Researchers had to wear protective suits withself-contained air supplies, they entered the lab through an airlock, it was like working in space. Theguidelines may have been draconian but at least they were clear and this gave the science a stamp ofapproval.

Cloning DNA in bacteriaPaul Bergimportance of being able to clone DNA using bacteria

You could take a colony and put it into a hundred-gallon vat and the bacteria would grow up and fillup the vat, and every cell in that vat would contain the piece of DNA that the original bacteriumpicked up when you mixed them with the DNA. So that showed you could clone DNA, and I thinkthat experiment is what galvanized the scientific community. It is in fact the experiment thatmotivated the moratorium letter, because it became clear you could put any kind of DNA into thatplasmid and get it into a bacterium, and so you could put toxin genes, you could put drug-resistantgenes, any kind of DNA you had access to could be put into a plasmid, put into a bacterium andcloned.

First recombinant DNAPaul Bergdescribing the first experiment with recombinant DNA

The first recombinant DNA made by using enzyme-created sticky ends, cohesion ends, cohesiveends, was actually done by my student, Janet Mertz. She took two different DNAs, each of them haddistinguishable properties, mixed them or cut them with the enzyme, mixed them, added an enzymethat could join ends to ends, and then showed she had created a molecule that now shared theproperties of the two starting materials. And the basic idea is that it's a very, very simple devicebased on really simple and very robust and ancient technology ofthe fountain pen. And the idea isthat the robot has a set of fountain pens that it dips into these microwells where each of these wellsrepresents a different human gene. The robot dips its fountain pens into these micro-wells and thenmoves over to a microscope slide and prints the tiny little drops of DNA. The grid you can see in thismicroarray slide is actually composed of 30,000 individual DNA dots, each targeted to match aspecific human gene.

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Cohesive ends & recombinationPaul Bergusing cohesive ends to make recombinant DNA molecules

Cohesive end in this particular context means that if you take two DNAs that have single strandsprotruding from their ends, and if these single strands are able to pair with each other by the samerules that DNA strands are held together, then these two molecules could come together. And whatJanet showed was that if two DNAs were cut with this particular enzyme, called EcoRI, then theycould be joined and fused together to make recombinant DNAs. Now that was a hugely importantdiscovery, because it bypassed the need for the complicated procedures that we had developed inorder to bring two molecules together.

Computing powerEwan Birneycomputational power of a processing farm

So we call this set-up a farm, there are 400 processors here and there are 400 computers and they'reall absolutely identically set up. The only thing that's different is their name -everything else iscompletely the same. So there's 400 processors, it's a very powerful processor chip: Alpha, CompaqAlpha chips. And quite a lot of memory, a gig's worth of memory in each box. So this is actually amassive amount of compute behind me. And what we do is we split our problem just into, say,20,000 pieces, and then each piece we send to one of these sort of worker bee nodes. And they dotheir work and then they give it back and then they say "I'm ready!", and they get another piece, andthey're just endlessly being driven by a master computer which is actually set somewhere else, thatdrives them, as you can see.

Reading the genomeEwan Birneyinterpreting the completed human genome sequence

All of human biology somehow connects back to the genome, and so anything that you can talk aboutin terms of human biology you can find some line that gets you back to some region of the genome.So just scanning over it is sort of wonderful. I was trained as a biochemist and as a molecularbiologist, and you just see, you know, thousands upon thousands of little stories about how your eyeswork or how your bones get put together or how the liver works and what happens here with thisparticular disease where, when this gene has a defect then this disease happens. And it's just a rich,massive story in some sense.

Impact of the genome projectsEwan Birneythe increased speed of gene searching

It used to take about, I don't know, eight to ten years to find a gene, say in the case of Huntington,there's this rather complicated process where they had to track many different families. And then

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there was this very annoying, painstaking process of them getting all the regions in test tubes, all thebits that they were looking at, and then carefully, experimentally checking each one. And it was eightto ten years, and a real pain in the arse. And these days you still have to gather all the families, butyou do not have to do this end-game. Now it's not five years or six years, it's three months orsomething like that. So the question now is not can I find the gene for Huntington, but why is it thata defect in Huntington gives you what we see, this disease? Why, what is going wrong, and, perhapsmore importantly what is this gene actually doing when it's going right? 'Cause that's the way weunderstand why in some cases, why it's going wrong. And so that's really what we've got the nexthundred years’ worth of molecular biology to do, is understanding the why, not the what any more.We've got the list and we now need to put together all the “why does this work, why does this givethis result?”

Walking down a chromosomeEwan Birneytraveling down the genome

So you imagine that you're traveling on the genome and, right at the start, there's this complicatedstructure right at the start which just effectively stops the DNA falling off the end of thechromosome. That's not quite true, but imagine that. I always imagine it as some sort of reallycomplicated regular structure that wraps around. And then suddenly you kick into the real genomeand you start marching down there, and a lot of it, if you looked at it, it would be like sort of trashbasically, you know, bits, shreds of, mainly of parasitic genes that have been jumping around ourgenome a while ago, and they've kind of got messed around and they make a real mess. So you haveto plow through some of this mess. And then you'll hit a gene and in fact, if you weren't a… if youdidn't know, you really wouldn't be able to tell the trash from the gene, but if you're in the know youcan say, aha, this is a gene. So you probably have a number of these regions going down the genome,and sometimes they're just one after the other after the other, and it's just straight, five, ten, fifteengenes all in a row. Quite why that is, we don't know. And then suddenly, you know, it's all chilled outand there's nothing going on, there's a sort of big gene that's just one gene in a region, and again wedon't know really why everything compresses in some regions and then spaces out in other regions.And then finally we get to these regions where we can't see anything at all. We see all the trash butit's just massive, it's a whole bacterial genome of nothing. And we call these deserts, and we still don'tknow what's going on in there, but the exciting thing is that if we, now we have the mouse genomeand we can look at the corresponding piece of mouse and the mouse has the desert as well. So hereare two things where we don't understand what's going on, but quite clearly humans and miceunderstand what's going on. There's probably something really exciting in there, we just need tounderstand it better.

Opposition to the HGPDavid Botsteinconcerns that the Human Genome Project marked the advent of "big biology"

Between the Santa Cruz and the Cold Spring Harbor meeting, there had been a lot of talk in and outof the press about doing the genome project. And there were very strong advocates whose views werewell reported and there was, also had generated a fair amount of opposition. And I must say I was

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among the opponents at that time. And the reason for it was that these advocates saw it as the adventof Big Biology, in the sense of Big Physics. And there were many reasons why many of us thoughtthat Big Biology in the sense of Big Physics was not appropriate or even a good idea.

Cost of the HGPDavid Botsteinassessing the cost of the Human Genome Project

Well, Wally Gilbert got up and made an estimate, which he'd also made in Santa Cruz, of how muchit would cost and that was three billion dollars. And there was good news and bad news about thethree billion dollars. The good news was that compared to the kind of money the physicists wereused to spending it was really sort of tissue paper, you know, for the bathrooms. It wasn't such a bigamount of money, and that made it a lot less scary. If you really could do it, and there was seriousdoubt as to whether you could do it for that kind of price, it didn't seem so distorted. It no longer wasthe Space Shuttle. The bad news was that three billion dollars was three billion dollars that no onehad planned on spending and one had to make a really good justification to everyone that this moneywould not be taken out of the hides of researchers, who after all were the intended recipients.

Locating disease genesDavid Botsteinlocating disease genes using markers

So the idea is that, for example, if a particular trait in corn lies next to a particular other trait, let'scall them A and B, and they're very close to each other on the same chromosome, then they areinherited together. So if you have green flowers and long shoots then when you get one or the othertrait, the other one comes along because they're inherited on the same bit of the same chromosome.Take a family in which you know Huntington is being inherited, take that family and look at all theparents and the offspring and see if you can find a correlation between a difference in the DNA thathas nothing to do with Huntington, but is just a polymorphism, just a difference, a spellingdifference is an analogy. So we said “Gee, what you could does take these Huntington families andjust get markers all over the genome that had this polymorphic, and then after the fact, just takingthe families as they walked into the clinic, look for that correlation.” And if we could have thatcorrelation, we would go from knowing that the Huntington disease is somewhere, to knowing thatit's somewhere on chromosome 5.

The impact of cloningHerb Boyerthe implications of cloning mammalian genes

One of the first things we realized when we did these experiments in the early 70s, was that now wecould isolate a piece of a mammalian gene or a fragment of a chromosome and you could reproduceit in large quantities, purify it, and it would be available to study. You could sequence it, you couldrearrange it and so on and so forth. And that was one of the first things that, that we realized whenwe had success with that experiment, that it was going to change understanding complex genomes.

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Birth of genetic engineeringHerb Boyersignificance of his experiment with Stanley Cohen to clone toad DNA

Cohen had spent years isolating genes from various species, until now he had nowhere to put them.Among his collection was a gene from an African clawed toad. In March 1973, Boyer and Cohenbegan an historic experiment to try to get the toad gene to work in a bacterium. Using Boyer’senzyme, Cohen sliced open the bacterium’s DNA and inserted the toad gene. But would thebacterium treat the new DNA as if it was its own? In another part of the bacterium they would findout. When the toad’s genetic instructions were fed through the cellular machinery, incredibly thebacteria were tricked into building pieces of toad. DNA’s instructions for creating living matter wereindeed universal. They could be transferred and read like files between computers. A singlebacterium multiplied rapidly into millions, each an identical copy of the original. Here was a livingfactory churning out fragments of toad, one life form manufacturing bits of another. Geneticengineering was born. It was a moment I’ll never forget. I could pretty much see that this was goingto change biological sciences. Yeah, well one of the things that was so revolutionary, I guess, aboutthis technology was that it was so easy when we actually did it.

Why study plasmids?Herb Boyerwhy Boyer and Stanley Cohen were interested in plasmids

We both had an interest in plasmids, these are small mini-chromosomes, usually non-essential forbacterial existence. But they had a strong medical importance because these plasmids carried genesthat gave bacteria resistance to antibiotics, so they have a big medical importance, and Stanley hadan interest in them. Also, because they were small pieces of DNA, they could be isolated and some oftheir properties could be manipulated and you could do experiments with them.

Code analogiesSydney Brennerthe idea of applying a code to DNA sequence

Many people have been interested in all of these ideas of codes and so on, and where did they allcome from? Now I think a lot of the thinking about a code was conditioned on thinking about theMorse Code, which those of us who had been Boy Scouts had to learn, you know, dot-dot-dot, dash-dash-dash, dot-dot-dot, SOS. So we thought of, really, a table and there'd be symbols on one sideand they would stand for what I think was more, was a wider thing because that's just the table that,which you go from the elements of one language into the elements of, if you like, a dictionary. Butthe idea of encoding information, that, I think had to come from the idea of computers.

Defining the geneSydney Brennermatching the gene to protein sequence

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I think the most important thing there was that immediately you could say, boy if we could find outhow the sequence of bases corresponds to the sequence of amino acids, because now we could definethe gene not just as a blob, not just as a bead on a string, but we could define the gene now as alength of DNA.

The coiled nature of DNASydney Brennerthe nature of DNA and RNA during protein synthesis

If you look at a bacterium and you say you've got these ribosomes and then you see these pictures inthe books of messenger RNA and the ribosomes running on messenger RNA, those are all wrong,those pictures are wrong. If you see these books you see a little DNA circle drawn in a bacterium,that's also wrong. The DNA is folded one thousand times. The length of DNA in a bacterium is abouta millimeter, the bacterium is one micrometer, one micron long. So the DNA is rolled up and foldedmany, many times inside this. And of course, when the messenger comes out, comes off the DNA,what you have to imagine is it threading itself through these almost stationary ribosomes, becausethey're big and they don't diffuse like a bunch of snakes. So if you really wanted to imagine thereality, it would be these snake-like molecules, writhing their way through this lattice of ribosomes ina bacterium.

Cell organizationSydney Brennerthe organization of cells in the body

Cells creep around, cells eat other cells, cells talk to other cells, cells listen to other cells. And the bestway I can think of, of communicating this is to just lean back and think about the town you live in,what is the machinery of the town? And what you know is that every morning we have all these littleunits called houses and out of these houses people come every morning and they travel andreassemble themselves into other units called banks or hospitals or factories. And it is those thingsthat exercise the functions of the city, whether they sell insurance or deposit checks or provide healthor education. And then every evening these units disassemble themselves and come back to thesefamily groups, and then they may assemble themselves in a different way, people go to parties, theygo to dances and so on. So what you have to have is a concept of all of that organization.

Explaining the Central DogmaSydney Brennerthe mechanism of protein synthesis and the virus (phage) experiment that proved it

So you can imagine what you have is, you have the DNA, there are the two strands of DNA, and thensomewhere here we had this ribosome which we'll draw as a box. And what we knew is that theinformation was coded in the sequence of bases here and what came out of the box was a protein,which is another amino acid sequence. And then, of course, the protein then folded up into thiselaborate three-dimensional structure. But the question is, how did the information get out of there,into the ribosome and then be translated into protein? Well the idea was, you'd copy a strand of this

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DNA, so we then get a strand, a single strand of RNA, that carries the message of the DNA, and that'swhy it's called messenger RNA, and it was taken into the ribosome here. And then here it wastranslated into the amino acid. So what you have to imagine then, if you have a bacterium where allof this is going in, we now inject a new DNA which comes out of this virus, which carries themessages for the viral proteins and then we knew there was this small RNA, this small amount ofRNA which was a copy of this chemically, I'd satisfied the base composition. And so what we had toprove is that this new RNA goes into pre-existing ribosomes, and then it reprograms them, so it stopsplaying rock and roll and now starts to play a little minuet of the phage, so that you can get the phageproteins made. And that was the experiment we set out to do.

The coding problemSydney Brennerthe problem posed by Watson and Crick’s model

Both Francis and Jim had seen that we immediately had what was, came to be called the codingproblem, in other words, how do these bases on the DNA specify the protein?

Gene expression patternsPat Browngene expression patterns in diseased cells

We knew that differences in the gene expression program were largely responsible for the differencesin the appearance and the behavior of the normal cells in our body, say a brain cell compared to aliver cell. And so it seemed natural that by looking systematically at the gene expression program incancer cells, we would be able to get a better understanding of why those cells are different fromtheir normal counterpart and why abreast cancer in one woman looks and behaves differently from abreast cancer in another woman.

Microarray analysis (includes Microarray animation)Pat Brownstudying gene expression using microarrays

And the basic idea is that it's a very, very simple device based on really simple and very robust andancient technology of the fountain pen. And the idea is that the robot has a set of fountain pens thatit dips into these microwells where each of these wells represents a different human gene. The robotdips its fountain pens into these micro-wells and then moves over to a microscope slide and printsthe tiny little drops of DNA. The grid you can see in this microarray slide is actually composed of30,000 individual DNA dots, each targeted to match a specific human gene. In order to work outwhich genes are being expressed, messenger RNA is first extracted from the cell sample and copiedback to DNA using an enzyme. This DNA, called cDNA, is complementary to the target gene, so willassociate with it, or hybridize with it, on the slide. After labeling with a fluorescent dye, the cDNA iswashed over the slide. The genes currently active in the cell can then be identified by the level of thefluorescence and the color of the spots.

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Germline therapyMario Capecchion needing to make germline therapy reversible

I mean the question is, you know, whether society is ready to think about things like, they're certainlyready to think about somatic gene therapy, I think that's more or less straight, I mean it's not easybut it is more like medicine as usual, because it only affects that individual. Now germ line genetherapy is more difficult to think about, simply because it not only affects the individual, but futuregenerations. So one of the things I would certainly mandate, if we even, ever consider doing germlinetherapy, is to make it reversible, that is to set up the system in such a way that if you take a cocktailthen you would be able to reverse all the changes you made. And this is very important because whatwe may think is brilliant today, twenty years from now may seem pretty stupid, and whatever theythink about in twenty years from now will be stupid twenty years hence. So it's very important tomake the system reversible.

Gene switchesMario Capecchiswitching genes on and off to study disease

There are techniques for being able to put a gene under a switch, so one way is to simply turn it offcompletely for all time with respect to the embryo, and that tells you all the places the gene is doingsomething. And, unfortunately, it may even kill the embryo and then you won't even be able to study,for example, what is this gene doing in the adult. So to obviate that problem, what we do is to put itunder a switch, and then we'll be able to turn it off in a particular space, for example the limb bud, orin the brain, and also at different times. We can look at it at different stages of embryogenesis, and infact we can wait and allow the gene to operate and make an adult and then turn it off and ask “whatis this gene doing in the adult?” As we speak, we have many genes that are operating all over ourbody, and this allows us to ask what is that gene doing, for example, in terms of processinginformation in our own brain. Most genes have multiple functions, are expressed in multiple tissues,and doing similar processes but in different tissues. And what we have to do is to be able to haveways, for example, of isolating the activity in a particular tissue, for example, the limb, or in thenervous system, depending on what we're particularly interested in studying.

Gene manipulationMario Capecchiusing embryonic stem cells to make mouse models

So in this case, we'll concentrate on this expression, where this gene is being used, what we call theexpression pattern, and we’ll neglect essentially this expression pattern which is throughout thewhole nervous system. So this machine, what it allows us to do is to…we place the ES cells, and theseare embryo-derived stem cells, so these are cells that are isolated from a very early embryo and theyare actually what are called pluripotent - they're capable of making all the parts of the body, forexample muscle, skin, nerve cells, teeth, whatever. And we can isolate them and maintain them inthis state of limbo where they're not committed and we can actually put them in the freezer, pullthem back out again, and it's in those cells that actually we do all of our genetic manipulations. And

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then once we've inactivated the gene in those ES cells, then we can take them and reintroduce themin an early embryo and there they get the signals to say, make bone, make heart, make skin, make thenervous system. And therefore that particular mutation that we created in a dish now is present in allthe tissues of the mouse.

Animal modelsMario Capecchiusing mouse models to study disease

So one of the advantages of working with mice is that in humans, if something's wrong with you, yougo to the doctor and then he can see what's happening. He'll say maybe you have cystic fibrosis, buthe can't go backwards in time and simply start studying the disease from the time you go. If wegenerate, for example, mice which have cystic fibrosis and which actually we have, then we canactually look retrospectively, we can make many, many mice that have just cystic fibrosis, we canlook at all stages of embryogenesis and then post-birth, and very carefully look at the pathology ofthat disease throughout time, and that way get a much greater detailed analysis of the disease. Andthen at the same time you can turn it around, and once you understand the disease in much greaterdetail, then you can actually start using the mouse as a model for developing new therapeutics ofthat, for that disease.

What came first: DNA or RNA?Tom Cechevidence that RNA evolved before DNA

Well, the question of how did life start is often referred to as a chicken and egg problem of earlyevolution: which came first, the DNA or the protein? After all, if you need an informational moleculein order to have inheritance, you need something, if not DNA, something functionally akin to DNA.But if nucleic acids are inert and cannot replicate themselves, they have to be copied by some activemachine. Well that's in modern cells typically a protein enzyme, so pretty improbable that at thebeginning of life you would have to have both the right nucleic acid and the protein that could copy itby random chemical processes both occurring in the same micro drop at the same moment. And apossible simplifying solution is that at the beginning there may have been RNA, ribonucleic acids,now that we know that RNA can both be informational and a catalyst, maybe RNA replicated itselfand then the proteins came on the scene a little bit later. Maybe the DNA was the last of thesemolecules to enter the scene, DNA being a more stable storage form of genetic information than RNAand maybe becomes important when you start getting a lot of chromosomes in an organism, youhave to maintain the fidelity of a lot of genetic information.

Definitions of lifeTom Cechlife, reproduction, and mutation

Well I've read what many philosophers and scientists have used for an origin of, for a definition ofwhat it is to be alive, and I think the simplest definition is that life is reproduction and mutation,

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interestingly. So if it were just a Xerox machine, making an exact replica, that's not quite sufficientfor life, there has to be some variation added with each round as well. Now of course that concept ofreproduction, and variation or mutation you might say is insufficient for life, but I think that'sembodied in everyone's definition. And then you can elaborate that and add additional requirementsthat a living entity has to somehow be separate, defined, encapsulated, enclosed, it has to be somekind of an entity separate from the rest of its environment, it has to be able to transduce energy.Those are, I think, the fundamental aspects of the definition of what is alive.

RNA splicingTom CechRNA is spliced

When we first got the copying of the DNA into RNA, what's called transcription, occurring in the testtube outside of the cell, we noticed that in addition to the gene being copied into RNA that the RNAwas undergoing splicing. Now this was… so the splicing being the cutting and rejoining reaction thatremoves or pops out the intron and produces the mature functional form of the RNA.

Patenting living organismsAnanda Chakrabartycreating and owning living organisms

A patent is not ownership as it is popularly known, a patent simply gives you a right to an inventionthat somebody else for a fixed period of time, let’s say 17 years to 20 years cannot use for commercialpurposes, for making money. I used to work for GE, GE had patents on toasters, refrigerators,washing machines, electrical circuits and so on. And that has never created any problem because weaccept the fact that inanimate objects can be patented. So that’s why when we came up with theliving microorganism as a patentable invention, there were a lot of people who thought that thatcouldn’t be done because that violates the fundamental laws of patenting, which is anything that’snatural cannot be patented.

The future of humansAnanda Chakrabartywhat will humans look like in 5,000 years?

So the question is … is there a limit to what the DNA could do? We are in the process of evolution, ifyou go back to even a few hundred thousand years, you know, there were no humans. So we haveevolved presumably from much lower forms of life and as we evolved our brain got bigger. We canthink much more clearly than in animals, or certainly lower forms of life and so on. Can you see thefuture and see how the human species would look like lets say 5,000 years from today will the brainbe even larger, will we really be able to think much more deeply, will we be able to travel betweenvarious planets and perhaps set up communication with other types of life forms that may or maynot involve DNA. Would be a very, very important question, I mean think if you leave it simply to myimagination I would say we are in the early stage of evolution, I think we are going to evolve, I thinkour brains would become larger perhaps even more efficient if it doesn’t increase too much in sizeand we’ll be able to do absolutely marvelous things in the future, so this is just the beginning.

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Chargaff’s ratiosErwin Chargaffthe DNA base ratio rules

We found, as I say, that there were constant quantities of each of the nucleotides in a given DNA,that is, if we isolated from three different sperm preparations human DNA, hydrolyzed theseseparately and determined quantitatively the content in the various nucleotides or bases, they wouldbe constant for the species but different for different species. So that, for instance, we could find the27 percent adenine, that means also 27 percent thymine, and in the other one we could find 52percent adenine and 52 percent thymine. So that a compound, these quantitative results wereconstant for the species but different from each other. So this is something which was completelyunusual, unknown in biology, I would say, that this, there are thousands of protein analyses and Idon't think one can find a single one in which certain amino acids are in identical characteristicamounts, that is in repeatable amounts. So that this was definitely a unique thing, for me at least.

The completed genomeBill Clintonthe completion of the draft human genome sequence

Ladies and gentlemen, the President of the United States accompanied by Dr. Francis Collins and Dr.J. Craig Venter. For a few happy hours on the morning of June the 26th, 2000, Celera and the publicconsortium forgot their differences and revealed their results together. Today the world is joining ushere in The East Room to celebrate the completion of the first survey of the entire human genome.Without a doubt this is the most important, most wondrous map ever produced by human kind. Ithink there was a great sense of relief that we'd made the story be the sequencing of the humangenome and the final sentence in my statement was "I'm happy that today the only race we aretalking about is the human race."

Cross-species recombinationStanley Cohenfirst experiment to recombine DNA from different species

The next step was to see if we could take DNA from a totally different biological source and to clonethat in E. coli. And for that we took a plasmid from Staphylococcus oreus and chopped that up intofragments using the EcoR1 enzyme, and ligated them to this pSC101 plasmid and selected for thepenicillin resistance that was present on the staphylococcal plasmid. And we showed at that timethat the E. coli plasmid could replicate, could propagate, and express DNA from a totally unrelatedbacterial species. And at that time it was very clear that this might be a generally applicable methodas we mentioned in our paper, the ability to clone DNA from an unrelated organism now opens up allsorts of opportunities for propagating foreign DNA in bacteria. And we proposed that even DNAfrom animal cells or plant cells might be propagated in bacteria using these methods.

More questions than answersFrancis Collinsapproaching population screening with caution

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Yeah, I've never tested myself for any of the genetic susceptibilities. I'd have to be convinced thatwe've arrived at the point where, for me, that information would have benefits that outshine thepotential risks of discrimination and other things. While we now offer cystic fibrosis carrier testing topeople of child-bearing age, my kids were already born by the time it was possible to do this, so thatwasn't so relevant for me. I've not gone ahead with other types of genetic testing for, say, Alzheimerdisease or diabetes because either the tests are rather imperfect in their prediction or because there'sno intervention available if you're at high risk. So, I'm basically following the recommendations thatthe genome effort has been making to everybody else, which is genetic tests are going to be extremelyvaluable. Most of them are not quite yet to the point where they ought to be available for generalpopulation screening, and before you take a test you need to know about the benefits and risks anddecide if it's right for you.

Science & faithFrancis Collinsreconciling working in science with faith in God

Well unlike, I suspect, many of my colleagues, I'm fairly comfortable speaking about the fact that avery important part of me, maybe the most important part of me is my faith in God, that is part ofwho I am. That's not something I grew up with, it's something I arrived at in my twenties by a seriesof sort of intellectual experiences, followed by a decision, and that's I think the way one has to cometo this. People often are puzzled by that, how, they ask, can you be looking at the sequence of thehuman genome which seems like the ultimate proof of Darwinism and the fact that this is not in anyway compatible with a divine intervention, and at the same time say you believe in God. Well I haveno trouble with that at all. Evolution is clearly the way in which all living organisms are related toeach other, I think that is unquestionable when you look at the data. At the same time, if God chosethe mechanism of evolution to create, and particularly to create creatures like ourselves who wouldhave some sense of desiring fellowship with God, who are we to say that that method, calledevolution, is a bad idea. It’s an incredibly elegant way to create. That puts me in the class of peoplecalled theistic evolutionists and most serious scientists I know, who are also people whose faith isimportant to them, fall into that same category because you see, if God has any meaning at all, God isoutside of what science can measure, God is also outside of time, so if God decided to use evolutionto create, he knew what the outcome was going to be, even if our methods of predicting it would saythat it was only going to be chance and you couldn't presume that. That assumes a certain boundingin time that doesn't apply to the supernatural and doesn't apply to God. So I find all of this incrediblysatisfying as a synthesis of what I know as a rigorous scientist and what I believe as a person of faith.And having both parts of those operate in a particular day in my life is very happy for me.

The challenge of gene huntingFrancis Collinsthe challenge of finding a disease gene

I like the analogy that trying to find a cause of a disease like cystic fibrosis or Huntington disease islike trying to find a single burned-out light bulb in the basement of a house somewhere in the UnitedStates. When you start out you don't even know what state to look in.

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The motivationFrancis Collinsthe justification for the Human Genome Project

So it took us nine years to find the cystic fibrosis gene, if you talk about all the time involved incollecting the families and doing all these steps, it took us ten years to find the Huntington diseasegene, a collaboration I had a chance to be part of as well. Took a little less than that to find theneurofibromatosis gene, but that's one where we had a very helpful clue that you don't always get.One of my conclusions from all of this is, we can't keep doing it this way, this is way too inefficient,the amount of wasted effort that comes along when you're feeling your way in the dark on every oneof these searches, is not going to be tolerable over the long run. And if we're really serious aboutapplying this same approach to diseases that are not inherited in such a straightforward way, likediabetes or heart disease or mental illness, we've got to have better tools. That was one of the reasonswhy, when the Human Genome Project was being talked about in the mid-1980s, I was its biggestfan, 'cause it was very clear that if one had that kind of set of tools, maps and sequences andtechnologies, that what had taken us so long could be telescoped into a much briefer interval.

DNA: the key to understandingFrancis Crickwhy the discovery of DNA's structure was so important

For a static structure, just a molecular structure to give such insight into all these, into even onefunction let alone a whole lot of other different…and such key functions because they are the keyfunctions of biology. So that’s why, essentially, it’s regarded as an important discovery because itcomplemented the original ideas of Darwin on evolution by natural selection plus genetics, whichhad started with Mendel, which would show that the genes were particulate and not blending. Andthen it showed what the molecular basis was, showed how the genes act, and in the last 10 or 15 yearshas given us a whole lot of new tools for fishing out genes, altering genes and so on which have gotimmense practical importance as well as being important theoretically.

The fascination of scienceFrancis Crickscience can give us a deeper understanding of the world

Now when you want to understand the world you have to go beyond those narrow limits up anddown, both in space and time. And then you find that there’s a uniformity and extraordinary thingshappening which you’d no idea of just looking at the world, and this is the fascination of sciencereally, I think, to uncover so much, which is not apparent just in everyday life.

Understanding the brainFrancis Crickdifficulties encountered when studying the human brain

What’s sometimes called reverse engineering, it happens in the commercial world when one firm

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produces a gadget and another firm buys it and tries to take it to pieces and find how it works. That’scalled reverse engineering, but in our case it’s reverse engineering in what you might call a foreignculture because we don’t understand the set of ideas which, as it were, there weren’t even any ideaswhich produced the brain. We don’t understand the principles which produced the brain.

DNA's deceptive "simplicity"Francis CrickDNA's processes that may be described simply are often quite complicated

Biological systems are the result of evolution and they produce very complicated things. Now thereason that DNA looks so beautiful and simple is it goes right back to near the origins of life wherethings had to be simple. But if you actually look at the actual process of DNA replication it isn’t at allthe way that we used to describe it as the sort of conceptual way, all sorts of funny things happen,you start off by making a bit of RNA, then you put the DNA on the end, then you cut the RNA out,then you fill it in all sorts of little things, you have to have proteins which will unwind the helix andnick it and then join it together again, so you get an enormously baroque complicated apparatuswhich actually does the job, which you could hardly say was simple and beautiful, it’s doing a lot ofsubsidiary things. It’s the basic idea, which is simple and beautiful.

The DMD approachKay Daviesgene replacement therapy in Duchenne muscular dystrophy (DMD)

The problem was because the disease was caused by the lack of the dystrophin gene, the only way toreplace it was either to replace it with dystrophin or to replace it with something that was verysimilar to it. And this looked like something very similar to it, and this was something that was ineverybody's cells, so we had a real opportunity of trying to make this gene replace dystrophin. Andthat's when of course we went ahead to try and prove that hypothesis. What was the sort of generaloutline on how you might use the related gene to cure muscular dystrophy? Well when we did thefirst experiment, we could see that it was there expressed in adult muscle. And therefore, if we couldpersuade the adult muscle to produce more of it, then by using chemical drugs in the normal drugtherapy fashion, then we might be able to induce more expression of eutrophin to alleviate thedisease.

Offering optionsKay Daviesimportance of choice regarding genetic testing

Well you have to give people choice. I mean, some people even these days would not choose to havepre-natal diagnosis for DMD, or even for Down. It has to be choice. And the other way round, if wedo have a genetic test for something, I think we should make it available to anybody that would wantit. We should not dictate to people what they want as families.

Genetic screeningKay Davies

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setting up a screen for muscular dystrophy

I was invited to give a lecture about the discovery of a gene that was very closely related, looked verysimilar to the gene that is missing in Duchenne muscular dystrophy [DMD]. And so I presented thatdata at the lecture at Cold Spring Harbor, and with the hypothesisthat if you could produce more ofthis in musclecells you might be able to cure the disease. And nobody really believed it, and Jimcame up to me afterwards and said that that was a real possibility and that he knew a company thatcould potentially help us do that next phase of the research. And therefore together with a starteramount of money from someone actually who was a parent of a patient in Cold Spring Harbor, wewent to see this company and we set up the screen.

A case for testingKay Daviestesting and managing genetic disorders

I think one of the positive things about genetic diagnosis, and Fragile X is a classic example of that, ifyou have a mentally-handicapped there's an enormous amount of guilt in the family, you know, wasthere an accident in pregnancy, you know, was I frightened by a horse, did I fall downstairs, did Idrink too much? If you give them a genetic diagnosis then they know it's fate, there's nothing thatthey could have done about it. So that's the first thing, so the guilt goes. And the second thing is ofcourse once you know what it is, you can manage it better. So we know FragileX patients haveparticular cognitive deficits, so there are teaching programs that we can design specifically for them.If they're hyperactive there are specific drugs that would be more suitable for a FragileX patient thananother sort of hyperactive child. So just the diagnosis makes a huge difference. What we don't wantto do is start labeling people and then them not being accepted into society, so what we want iseffective management so these children can actually be more acceptable in normal schools, ratherthan being told this is just a behaviorally-disturbed child who doesn't learn very well. And that Ithink is how I would like to look at it.

Developing other cancer drugsBrian Drukerapplying the Gleevec model to other cancers

If you could understand what drives the growth of cancer, target that with a specific treatment that’seffective and non-toxic, you can do that with every single cancer. And what's unique about chronicmyeloid leukemia is not the cancer, but our understanding of it. We understand what drives thegrowth and we can treat it early in the course of the disease. And so many people say well, if it’sadvanced cancer it’s gonna be more difficult. And I say I agree. Advanced cancers are difficult, butwhat about early cancers? What if we can identify cancers in the earlier stages, identify what drivestheir growth and shut them down early? So we still have a lot of work in identifying cancers early andidentifying what distinguishes them from normal. But that’s the paradigm. Identify them early,identify what's wrong early and treat them before they're an advanced malignancy. That’s theGleevec paradigm, and if we can do that with every single cancer then there’ll be a before Gleevecand after Gleevec.

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Leukemia: the Gleevec storyBrian Drukerthe development of Gleevec, a drug to treat leukemia

So the Gleevec targets a protein called BCR/Abl and this BCR/Abl protein belongs to a family ofenzymes that regulate cell growth. It comes about because of two pieces of DNA exchangingsegments. So it’s a genetic disorder, it’s something that people pick up, it’s not inherited, it’ssomething people pick up in their bone marrow during their lifetime in one blood cell. Two pieces ofDNA rearranged create this abnormal protein, it’s sort of like an accelerator of a car getting stuck on,telling cells to grow continuously and they continue to grow and divide causing leukemia. So it’sdirectly related to DNA. What we've done…DNA as you know makes RNA makes protein, and it’s thisprotein that drives the growth of the leukemia cells. We shut down this protein, we shut down thisleukemia, and when these cells die, the genetic abnormality in those cells goes away too.

Explaining life through scienceRosalind Franklin’s sisterreading a letter from Rosalind to her father

Science for me gives a partial explanation of life. In so far as it goes it is based on fact, experience,and experiment. Your theories are those which you and many other people find easiest andpleasantest to believe but as far as I can see they have no foundation other than that they lead to apleasant view of life and an exaggerated view of our own importance. Anyone able to believe in allthat religion implies obviously must have such faith but I maintain that faith in this world is perfectlypossible without faith in another world.

Experiments to find RNAWalter Gilbertexperiments carried out to isolate RNA

The three of us, Jim, François and I, would do an experiment together, François generally holding agigantic flask full of bacteria and shaking it violently, I holding a bottle of radioactive phosphorous,20 millicuries, Jim holding a stopwatch, and then Jim would push the button, shout “Go.” I wouldpour the radioactive phosphorous into this big flask, François would shake it like mad, Jim wouldshout “Stop!” Then end of twenty seconds, and we'd pour this flask of radioactive material over a bigfifty gallon can full of ice and try to stop the growth of bacteria immediately, hopefully not splashingtoo much radioactive material around the room. We'd then spend hours harvesting the bacteria,spinning in a centrifuge down to collect the bacteria out of this large mass of radioactive material,put the radioactive material away to decay and not harm anybody, and then try to analyze thebacteria to see if we could see an RNA molecule labeled by this radioactive phosphorous in this veryshort pulse. And we did experiments in that throughout the summer. And from, starting in the earlypart of the summer we had evidence that there was such an intermediate and we published a paperon that around December of that year.

Gene regulation

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Walter Gilbertthe regulation of genes by control proteins

I and Benno Müller-Hill, who was at that time a post-doc with me, started to work on that probleminvolving the repressor for the lactose operon, and on a set of genes involving the metabolism of thesugar lactose. We worked on it because we knew it was a fundamental problem in biology, and theproblem that we were attacking was what is the nature of the product made by one gene that turnsoff a second one. The answer in those days that we came to was the product is a protein that goes andbinds to DNA and prevents a second gene from functioning. The small molecule of sugar coming intothe cell inactivates that protein,makes it release its hold on DNA, and then the genes begin tofunction. There are only ten molecules of that protein in a bacterial cell. It was extremely hard toidentify, and we did a set of experiments in the middle Sixties that actually enabled us to identify thatprotein, it was the first control protein identified.

The repressor/inducer systemWalter Gilbertusing bacterial mutants to study molecular interactions

We knew that the molecule should interact with the sugar that was coming into the cell and so wecould use a radioactive form of that sugar to try to look for a molecule that it interacted with insidethe bacterial cell. We knew that that interaction was, we knew something about the strength that, ofthat interaction and we could actually decide that that interaction was probably too weak for us to beable to see it. So Benno Müller-Hill modified the bacteria, made a mutation, a genetic mutation, thatchanged the way in which the repressor bound to the small molecule we call an inducer, and made amutation so the repressor now worked more tightly, bound to the inducer at a lower concentration.And then we took those bacteria, and looked for the interaction between the, a protein and the smallmolecule, by using a radioactive small molecule, and asking, actually, we put the protein inside alittle sac that the small molecule could move in and out of, could we find an excess of moleculesbound inside the sac because they were attached to the protein. And we actually made thatexperiment work.

Insulin productionDave Goeddelthe Genentech method of producing human insulin

Okay the, the Genentech approach to the insulin project was to ultimately create a gene that could berecognized by bacteria and produce human insulin. We had three major steps to do that, that is howwe divided the program. The first step was the chemical synthesis of the gene itself, and assemblingit; the second step was to take the chemically synthesized gene and to clone it and by that meansinsert it into bacteria in a way that the gene can be replicated in, producing unlimited quantities. Andthe third step is the production or manufacturing step, which is really coaxing the bacteria intorecognizing this sequence and actually producing the final product, the human insulin.

Influencing our evolutionRaymond Gosling

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eliminating faults in our genetic programming

People will say oh well, the way to look at it is, are we behaving in an ethical fashion. People nowalready say, oh it's great, we will be able to use gene therapy to prevent human beings from havingnasty things like cystic fibrosis or diabetes and you can go on and on, and we can eliminate that. Wellif you can do that, the next thing of course is designer babies. And so you will be able to influence ourrate of evolution and our direction of evolution as has never happened before. Evolution has, inDarwinian principle been influenced by the environment and the survival of the fittest and so on.However, mankind has already mucked that up, we've already interfered in that in one way, in thesense that we have enabled human beings who once would have died of various disorders which wenow know to be due to genetic program, to, if you like, faults in the genetic programming, which wecan now eliminate. But we've enabled them to live long enough to reproduce, so we've already badlypolluted the human gene pool. Now if we've done that, would it be ethical to leave it polluted, wouldit not be just as ethical now to kind of redress the balance.

An earlier DNA modelRaymond GoslingFranklin’s analysis of Watson and Crick’s early model of DNA

They built their model with the phosphoruses on the inside, giving… because they would sit oneabove the other and give a nice backbone if you like, like a human being, and then all the bases stuckout like ribs. And so she laughed at them, much to their discomfiture I think, and said “Oh look,you've got it inside out, the phosphoruses are all on the outside and they are, for the followingreasons.” And she explained her experimental work And her reasoning and so they sat back andabsorbed that and, suitably chastened, they… I don't know what they did. They probably didn't doanything for a while. Her view was you could build models all day but how did you prove which onewas right? On the other hand, if you made the measurements, you did all the corrective geometryand you put them into the equations, you would let the data speak for itself. And out of that willcome a definitive structure.

Clue: X-ray diffractionRaymond Goslinghow the X-ray diffraction camera worksMaurice Wilkinsthe X-ray diffraction picture that revealed the helix

You're looking at the pattern produced in this camera by shining a beam of X-rays through a singlefiber. So you have a micro-camera there which has a lead glass collimator which is about as thick asmy finger and about that long, with a hundred m [micron] hole drilled down the middle. It's simply apiece of capillary tubing but it's made of lead glass, so all the X-rays are absorbed except those thatgo straight down the hole. And if you line that up here in this, so it's in line with the X-ray beam, youthen have a film set at the back of this camera. The camera is filled with hydrogen, the X-ray beamgoes through the fiber and then the diffracted beams flash out in discrete copies of the incident beamin width, but of course the direction is different, and they're intercepted by a piece of film and thatfilm gives rise to all these spots. So that is the pattern you see when you develop the photographic

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negative - it's been blackened by those diffracted beams, and you can see there is a regular pattern tothat, and that regularity is a result of the regular pattern of the crystals of DNA in the fiber. It wasvery exciting to see all the spots on the photographic film which – fairly sharp spots – and it showedthis sort of X-type of OXO type of cross-pattern which was an indication of a helix.

An elegant structureRaymond Goslingthe elegant simplicity of Watson and Crick’s model

Having got the specific pairing he couldn't wait to put it all together. And of course it went togetherand then when they'd done it they must have been absolutely awestruck in its elegant simplicity. Andthey realized that they'd explained the secret of cell reproduction, the secret of life if you, if you will.

Race to discover the structureRaymond Goslingthe race between King’s College, London, and other groups to define the structure of DNA

Three teams, three different approaches and Watson and Crick weren't the favorites. The race, Ithink, as Maurice Wilkins saw it, was principally Linus Pauling and King's [College]. He didn't realizethat Crick and Watson were working on this. They'd been told by Lawrence Bragg not to. It was, Ibelieve, very much James Watson who had that sense of wonder, which, as I say, gave him the driveand the focus. But at King's, if anybody did perhaps feel it, Wilkins did, because he often said to me,you know, we should all wake up, we should all try a number of different things instead of ploddingalong trying to solve the X-ray diffraction pattern, because we are in a race. It is an importantproblem and there are other people, I mean there was this bogeyman sitting in America called LinusPauling who already had two Nobel Prizes, not one.

Clue: position of phosphatesRaymond Goslingrealizing phosphates are on the outside of the structure

Because we'd been working away and we knew from the way we could take water in and out of thestructure and further change the structure from one crystalline to a paracrystalline thing with ease,that the things that the water were attaching to was the sodium and phosphorus. You had thephosphate group and the sodiums sticking to that, and that attracted a whole lot of water moleculesaround it. So the water going in and out meant that the phosphoruses must be on the outside.

Tracking human historyMichael F. Hammerusing the Y chromosome and other genomic regions to track human history

Although we can trace these chromosomes back to single ancestors in the past, there are manypeople living in the past. It’s only through sort of the lottery of who has children and who doesn’t,over many generations, that some lineages are lost from the population. And so we don’t see themtoday but those people lived and made contributions and may have made contributions in their

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autosomal DNA, their X-chromosome DNA, their mitochondrial DNA that we’re not seeing bylooking at Y chromosomes. Each gene, part of the genome has got its own genealogical history thatgoes back to maybe different people. So we can trace the Y chromosome back to a single ancestor andmay call that Y-chromosome “Adam” but that man did not contribute his genes, necessarily othergenes, to the population living today. So there are many stories being told by tracing different genesin the genome back to common ancestors. Because of the process of recombination some of thosestories are too complicated to unravel because each gene is kind of a mosaic of many histories.

The evolutionary puzzleMichael F. Hammergenetic data must be part of a framework

From genetics alone we can’t tell all that much. We need to have a context to work in. So, if I’minterested in the peopling of the Americas – how long ago did people move into the Americas? Howmany people moved into the Americas? How many times did they move in the Americas? – I can getgenetic data that will show me patterns of variation in the Americas and I can compare those datawith patterns of variation in Asia. But I need calibration points from the archaeological record, toknow when we see evidence of culture in the Americas. How does that culture relate to culture inAsia? And it’s a comparative process through genetics and archeology, and sometimes linguistics andsometimes the fossil record, if we’re going back deeper in time. You have to put the picture togetherwith all of those pieces of the puzzle. One piece of the puzzle alone won’t give you the whole picture.So we shouldn’t lose sight of that. As powerful as genetics is, as a tool, to look at our own history, itcan’t tell us anything by itself. It has to be in a comparison framework.

Harrington familyRoby Harringtonhis relationship with his son who has Down syndrome

On Sunday he and I go to the beach and we've been doing this for four or five years and he and I goout into the water and if it’s a calm day I hold his hand, if it’s a slightly wavier day I hold him and youknow he holds real tight and we jump over the waves and then he goes in and sits on the shore and Iride a few waves and he likes to grade me but, two weeks ago, three weeks ago, on our last trip outthis fall, we went into the water, it was a pretty calm day we were about up to our chest, my chest, Iwas holding him and all of a sudden he asked to be let go and I let him go, he's become quitecomfortable in the water and he rode this modest wave and probably went seven feet and he turnedover and then popped up, startled at first and then he realized that he had ridden a wave all byhimself and the look on his face and the look on my face and he just opened his arms and it was oneof those moments of just such pure joy that he and I had anticipated this for four years now herealized he had achieved this and I realized I had achieved it and I didn't think is he doing this beforeother kids do it his age, it was just the accomplishment that we shared together that I think is just, itssomething that I have with him that I wish more people that were considering life choices could feelthat, could see that, could realize that yes the hurdles, there's a longer approach and you approach ittogether, he clears them by himself but there is just a magic in watching him do it, that as your otherkids zoom through their hurdles you might not appreciate in all the same ways.

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Sequencing DNALeroy Hoodinside a DNA sequencing machine

The individual samples are placed in the wells at the top of the gel, the electrophoretic field is turnedon and each of the samples migrate down the gel, small bands traveling more rapidly than largebands and at the bottom of the gel is a laser that scans across each of the lanes, reading out the colorof their four different bands, red, green, yellow and blue, and those represent the four different DNAdyes. The information is recorded in a computer and then the biologist has access to interpreting thedata after the run is finished.

The digital codeLeroy HoodDNA is a digital code that can be read

Well, if you think about the most important observation that came out of Watson and Crick'sstructure of DNA, it is that DNA is a digital code, a code that's embedded on 24 digital strings in thehuman. And the idea that you could actually read out the digital information that was the very coresource code for human development was an enormously exciting idea.

Implications for the futureLeroy Hoodnew tools for redesigning life

A guess that I would venture forth would, perhaps we could do an entire human sequence in a fewdays for a cost of say ten thousand dollars, you know, in contrast with what the first human genomewas, it took perhaps a year and certainly cost two or three hundred million dollars. Then we can startto redesign creatures to do things in very rational ways, not by kind of the classic hit and missexperiments that we have to carry out today. But for example, you might think about designingwheat that could do five things wheat can't do today, give us four nutrients plus make the food valuemuch higher than it is now, for example. So once we understand the regulatory circuitry and once wehave a kind of a lexicon, a parts list of what are the fundamental sub circuits, then we can begin tothink about redesigning life in quite striking ways.

Future of medicineLeroy Hoodpredictive and preventative personalized medicine

My prediction is, in the next ten to fifteen years we will have identified hundreds of genes that play amajor role in predisposing to cardiovascular disease, cancer, autoimmune disease, metabolic disease,neurological diseases. And what we'll actually be able to do at a relatively early period of your life istake a sample of your blood, analyze your genes, put those in a computer and we can write out aprobabilistic health history of what is likely to happen to you. And of course it's an anathema formedicine to be able to do prediction without prevention, so what we will have to follow that with isusing the systems approaches I've talked about to place these defective genes in the networks within

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which they operate. And in doing so we'll learn a lot of ways to circumvent their limitations, whetherit's drugs, whether it's embryonic stem cell therapy, whether it's germline gene therapy, there'll be awhole variety of very powerful preventive measures that one can take, such that after you're giventhis probabilistic history you can take these pills and you won't have to worry about kidney disease orcardiovascular disease. And that will of course be the predictive on the one hand and the preventivemedicine on the other hand. And what the two of them add up to in a very interesting way, ispersonalized medicine.

Redesigning organismsLeroy Hoodtechnological advances may allow us to redesign life

What is exciting about this is, the core of understanding the essence of biology is having many ofthese digital source codes for many, many different kinds of creatures, because by being able tocompare these different source codes, we can not only deduce the logic of how genes work, but thelogic of how regulatory networks work. And the really exciting idea is, once we begin to understandhow regulatory networks work, then we can start to redesign creatures to do things in very rationalways, not by kind of the classic hit and miss experiments that we have to carry out today. But forexample, you might think about designing wheat that could do five things wheat can't do today, giveus four nutrients plus make the food value much higher than it is now, for example. So once weunderstand the regulatory circuitry and once we have a kind of a lexicon, a parts list of what are thefundamental sub-circuits, then we can begin to think about redesigning life in quite striking ways.

Regulatory networksLeroy Hoodthe importance of regulatory systems for evolution

To give you an example of how important regulation is, if we compare the genomes of the human andchimps, we note they differ by about one letter in a hundred, and in fact that probably changes mostof their genes very little at all. But if you look at the brain of a human, and the brain of a chimp, yousee that the human brain is bigger and far more convoluted and has a much denser concentration ofnerve cells. So it means the regulatory machinery that has developed a brain has changed incrediblyin just a period of six million years or so when the two species diverged from one another. So whatthat says quite clearly is, it's the regulatory networks that in a deep sense explain development andphysiology and even evolution in many interesting ways. And that's why they are such an importantpart of systems biology.

Analyzing your genesLeroy Hoodunderstanding the genome will lead to medical advances

What will be useful in the future is to be able to look at the variability that correlates with disease.And my prediction is, in the next ten to fifteen years we will have identified hundreds of genes thatplay a major role in predisposing to cardiovascular disease, cancer, auto-immune disease, metabolicdiseases, neurologic diseases. And what we'll actually be able to do at a relatively early period of your

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life is take a sample of your blood, analyze your genes, put those in a computer and we can write outa probabilistic health history of what is likely to happen to you.

Manipulating living systemsRobert Horschmanufacturing new proteins in living systems

In two billion years of evolution just the bare surface has been scratched of what's geneticallypossible with the genetic code in DNA. And we're seeing the possibility to use DNA for computationsystems; we're seeing the ability to start producing starches and plastics in green plants instead ofcement and steel plants. We're seeing the possibility to use proteins in medicine that are grown ingreen plants instead of fermented in cement and steel plants. Just all kinds of functionality andthings of value based on improving the information content of genes and living systems that cannotbe done with fossil fuels or mining.

DNA transfer: gene gunRobert Horschinserting genes into plant cells using a gene gun

The major alternative to the use of agrobacterium to get genes into living plant cells is a physicalmethod called the gene gun, which literally shoots gold or tungsten particles into living plant cells,carrying DNA on their surface. The first embodiment of this by the fellow at Cornell, John Sanford,whoinvented it, actually used a .22 caliber blank to accelerate a plastic bullet with a drop of tungstenand DNA on the tip towards plant cells. The plastic bullet would hit a doughnut-shaped plate with ahole in the middle and the tungsten would spray out the other end but the plastic bullet would stop,and the plant cells in the bottom would be bombarded at gunshot velocities. It turned out it worked.

First transgenic cropRobert Horschthe first transgenic crop, engineered by Monsanto

On June 2nd, 1987 an historic occasion occurred in this field. It was early in the morning when we leftChesterfield Research Center in anticipation of planting the first field test of genetically-engineeredtomatoes with one of three different traits, either herbicide resistance, virus resistance or insectresistance. In probably the most important sense, the field trial was a huge success, everything wasdone according to the protocol, the plants grew well and we were able to get a very good scientificassessment of the performance of each new trait that had been added to the tomatoes. Unfortunatelythat performance, while we were able to scientifically assess it very well, just didn't measure up towhat a farmer would expect in a crop, in a trait that they would buy and use on their farm. But thatdata set was essential to go back to the lab and make the next generation improvements in each ofthose traits. The farm here at Jerseyville has become one of our main centers for continuing todevelop new traits and combinations of traits in our core crops of corn and soybean in particular. Forexample, we have developed and tested over several years our newest product, which we hope to

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have out in the summer of 2003, a gene that protects corn against the most serious pest in theUnited States, the corn rootworm.

DNA transfer: agrobacteriumRobert Horschtransferring genes into plant cells using agrobacterium

Well once we had the engineered gene, the selectable marker, and had it put in a vector in theagrobacterium, the rest was the cell culture and selection. So we would start with a bleach-treatedpetunia leaf that the bleach kills the surface micro-organisms that might contaminate things, andthen we use actually a common paper punch that's been soaked in alcohol to clean it up, and punchdiscs out of the leaf. These discs then are nice uniform size pieces of the leaf tissue and have awounded edge all the way around them. The agrobacterium grown in a simple broth then could beinoculated into that wounded edge and give a uniform infiltration of the bacterium with the woundedplant cells. After about two days of this co-cultivation, you could use an antibiotic that would kill theagrobacterium, because it had already transferred its DNA from itself into the plant genome. Andthen you could start selecting for the selectable marker and stimulating the regrowth of individualplant cells in the edge of the leaf disc to grow back to a whole plant.

Sequencing genomesMike Hunkapillerthe speed of sequencing since automation

The first free-living organism was sequenced at The Institute for Genomic Research in 1995, it was afew million base pairs long. It took them about six to nine months to collect the data and interpret itand get a close sequence. If you jump forward a little bit to three years later, the first complexorganism was done, Drosophila, in about the same amount of time -it was a hundred and twentymillion base pairs long. You jump forward another year and you have the human sequence done atthree billion base pairs, also in about nine months. And so just over the sort of mid-life evolution ofthe automated technology, you had far more than a thousand-fold increase in throughput.

The lac operon modelFrançois Jacobhis model for bacterial gene regulation

So, by comparing the system of the viruses, the lysogenic bacteria on the one hand and the inducedsynthesis of galactosidase, it was clear that it was two very similar systems, and by comparing thissystem we came to a model which I am trying to describe now, the idea what you had a genecontrolling the structure of proteins which we called structural gene, and we can call that SG1, SG2,cloned to each other, for instance the Z gene of galactosidase and the Y gene permease. Andsomewhere else which could be either far or close to the other, a regulatory gene, that is a gene whichcontrols the expression of these two structural genes. So the idea was that from this you had amessenger, that the gene which is it's DNA, a messenger which was RNA, and from this messenger

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proteins were done. So that goes like that. Now the regulation was by what we call this negativesystem, that is a regulatory gene made a product which at first we thought was RNA but finally wasprotein, so it made a protein which we called a repressor. And this repressor was acting on a smallsequence of DNA adjacent to the structure which we called the operator. So there was here… so theregulatory gene produced a repressor, a protein, which binds to a specific sequence which is theoperator, which control the initiation of messenger production. And the lactose, that is the inducer,was supposed to interact with the repressor and prevent him from attaching to the operator.*So therepressor was a double sided structure, one which recognizes the operator and the other whichrecognizes the small molecule, the lactose. And this we call the negative system because it blocked, sothat was the model which we call operon.

A pre-1953 notionFrançois Jacobbiology prior to discovery of the double helix

I wanted to say when I started the biology, in the fifties, the idea was that the molecules from oneorganism were very different from the molecules of another organism. For instance, cows had cowmolecules and goats had goat molecules and snakes had snake molecules, and it was because theywere made of cow molecules that a cow was a cow. And then it turned out a little bit later that certainmolecules were very similar from one species to another, for instance hemoglobin, that thehemoglobin of a horse and a cow or a human turned out to be very similar. And progressively as itwas possible to know about the primary structure of the sequence of protein molecule, it turned outthat more and more of molecules of various organisms were very similar.

Manic depressionKay Jamisonmental illness and complicated choices

If I were asked, would I choose to have this illness, with a huge caveat that I have a treatable form ofthis illness, a very treatable form of the illness, and most forms of the illness are treatable, but Irespond extremely well to lithium so that's a caveat. If I didn't respond well, I would have a totallydifferent answer. But because I do respond well to treatment I would say I would choose to have thisillness, it is who I am, it is built into my genes, I mean it is my temperament, it is my personality, itis, everything that I know to be true about myself is predicated to some extent on having beeninfluenced by this illness, good, bad, and indifferent. But would I choose not to have gone throughthe suffering and all the years of pain and the lost years of which there are many, many just lost yearsfrom my life, I wouldn't go through that again, I wouldn't inflict that on anybody I know, I wouldn'tbegin to think of inflicting it on anybody. So it's complicated.

Testing for a reasonKay Jamisonschizophrenia: a case for testing

Well first of all, I mean first of all schizophrenia's a very good example, because if you'dasked thisquestion ten years ago I think most of us who are in psychology or psychiatry would say, you know,

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wow, this is a real exception.This is such a terrible, dreadful, degenerative disease that it's not, youknow, whatwould the options be, would you choose? But if you look at the treatments that havecomeout in the last ten years for schizophrenia, and the options that people have, and ifyou think aboutwhat's going to happen evenin twenty years from now with schizophrenia, I think it's going to be verydifferent. I think people will correct it, the problem of what causes it, and I think there will be muchbetter treatments. So people with schizophrenia now have very different lives than ten years ago, so Ithink it actually makes the case.

Vanilla childrenKay Jamisonfor diversity and against narrowing the options

I don't take the views of, that you don't reproduce under those circumstances, I find that a chillingthought. And some of it I'm sure is from a very personal point of view I find it chilling, but I also as aprofessional and as a clinician and as a scientist and as a member of society I find it a chillingthought. I love the fact that we are as diverse a species as we are, I love the fact that we have such adiversity of temperament and such a diversity of intellectual styles and ways of seeing the world andways of going through the world. And I don't want to start narrowing that range, you know, I think Iwant to give people a lot of options in terms of treatment and a lot of options about what they dowith their lives. I don't want to give people a sense that they have to start narrowing and narrowingand narrowing and producing kind of little “vanilla children,” it's a kind of a ghastly thought.

Mental illness & creativityKay Jamisongenetic links between mental illness and creativity

One thing that is clear is that, if you look at the scientific literature, there are more than twentystudies showing a very much elevated rate of depression and manic depression in people who arehighly creative. Obviously most people who are highly creative are… don't suffer from any kind ofpsychopathology, they aren't particularly moody, they are… don't suffer from depression. It's ratherthat there is a disproportionate rate. And I think that when you're talking about genius, when you'retalking about someone like, with the mind of someone like [Lord George] Byron, for example, orVirginia Woolf, I think you're talking about people who are by definition…it's a very unusual set ofcircumstances to, that leads to that kind of mind and ability. And in both of those instances, a verystrong family history, it's a genetic illness, a very strong family history of depression and mania andsuicide. And in both of thoseinstances you could see how these illnesses both gave them a range ofexperiencefrom which they wrote, but it also gave an intensity and a power and a fluency tothatwhich they wrote. Would they have been thatway anyway, without the illness? Who knows?

Protecting diversityKay Jamisonmanic depressives: an endangered but valued species

My concern would be that at some point manic depressives would become an endangered speciesand that they would not have their representation as an intellectual force in their own right and

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energy force in their own right, a way of acting in their own right. And again, it's not to romanticize abad disease, because I'm not doing that, but I think that it's complicated and I think you want torespect how complicated it is, that it's not just a severe mental illness, the severe mental illness isattached to temperament, and it's attached to cognitive style and it's attached to many wonderfulattributes in terms of exuberance and high energy levels and high-voltage people. And so you don'twant to risk trimming those folks too much, all the while you want to, the great advantage of geneticsthat you would be able to diagnose it more accurately and earlier and get much more specific andmeaningful treatments. And those are the wonderful aspects of it. The possible down side would bethat you would be giving mankind the power to eliminate something that might be potentially veryuseful as long as it's treatable.

DNA fingerprintingAlec Jeffreysusing minisatellites (tandem DNA repeats) to create unique genetic profiles

We discovered this shared sequence motif, shared by these different minisatellite repeats, so wethought okay, what we need to do now if we're going to detect lots of these really good minisatellites,is you take this shared motif, make it as a tandem repeat, and use that as a probe, that should be verygood at detecting minisatellites. So I think it was mid-September, did the experiment, I think it wason a Friday, left the X-ray film exposing and the result came off on Monday morning, I think it wasabout nine o'clock in the morning, where I just went into the dark-room, we'd done this experimenton a, what we call a blot, just a little membrane containing various DNA samples. It was prettymessy, it was very, very messy but it was clearly working. What we were detecting were lots of bits ofDNA and in the humans at least they were clearly variable, very variable, and in fact what we'd gotwas our first very murky DNA fingerprint. And at that point the penny dropped I thought wait aminute, we've actually solved a totally different problem here, in theory. If these patterns, if we canimprove this technology, these patterns are going to look as if they're going to be extremelyindividual-specific. And not only that, we can begin to see from our little family group that they seemto be rather simply inherited as well, so as if a child was made up of half of mum's bands and half ofdad's bands on this DNA fingerprint. So I thought okay, we've got a possibility here of using this foridentification, so we immediately thought forensics; possibility here maybe we could use this forestablishing family relationships, so immediately started thinking of paternity disputes. We alsothought about other potential applications and these are all thoughts that were coming in, in a veryembryonic form into my mind, literally within the first five minutes of actually peering at the bit ofX-ray film.

GM crop concernsJim Kentraising concerns associated with GM crop production

In the U.S. when a crop is grown it tends to be grown in a monoculture where you have thousandsand hundreds of thousands of acres being planted with the same crop. And so basically it's possiblefor a company to sell seed to farmers and, farmers have been tending, in the US at least, to be buyingtheir seed ever since the hybrid revolution of the Seventies, rather than just getting it from theprevious year's crop. And so the net effect then is that when something is put into a serial genome it

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can go from being tested in the lab, to all of a sudden it's covering a third of the acreage in the U.S.And I just, that scares me a little bit, I wish that it would scale up much more slowly to have a chanceto sort of see where the problems may be.

An important gene clusterJim Kenta cluster of immunity genes on chromosome six

You'll often see like clusters of genes. A very important one is, for instance, on chromosome 6.There's a section, the major histocompatibility locus we call it. And this is an area that is, actually it'skind of the heart of your immune system all clustered together in this relatively dense area onchromosome 6.

Coding vs. non-codingJim Kenthow much of the genome is active?

It's not clear how much of the genome is doing anything. We know that about one percent of it seemsto be involved with actively producing proteins, the sort of, the moving parts of the cell. But thenwhat the rest does is fairly mysterious. We get a little bit of a handle on this by comparing it with themouse, and from mouse comparisons and seeing what's conserved between mouse and human, itlooks like, you know, more, perhaps five percent is serving some function. And some of that is thingsthat we know about, some of it is bits involved with turning genes on and off, rather than the actualgenes themselves.

Assembling the fragmentsJim Kentproblems assembling the genome fragments

Actually what makes DNA assembly maybe more challenging even than Russian prose is that it canactually be read forward and backwards, and so you have to see if it goes this way, and ifit doesn't gothis way, maybe it goes that way. And this bit actually was particularly hard for me because I had alittle bit of dyslexia when I was a kid and I was always getting my letters reversed. And so sort ofhaving, does it go this way or that way and, I don't know, it was kind of confusing, that I would all thetime be getting things backwards and would just have to proceed by trial and error. Fortunately therewould be only two cases to do, you know, if it wasn't this way then we’ll just swap the sign and, andmaybe it'll work that way. Then another thing that makes DNA assembly a challenge is that thehuman genome is actually in effect, 'fact it's not really like prose and in some ways it's more likepoetry or a song even, in that it has a lot of repeating elements. And so you can imagine like a, sayyou were trying to assemble “Mary Had a Little Lamb” from pieces, you'd have "Mary had a littlelamb, little lamb, little lamb, Mary had a little lamb whose fleece was white as snow” okay. Well thenif you, if you just had the phrase "little lamb" where are you going to put that in this song? You canput it here, you could put it here, you could put it here, you just don't know. And, it's actually thesingle thing that makes the assembly difficult is coping with the repeats.

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Random mutationsJim Kentrandom mutations are necessary for evolution

It turns out you can learn a lot by comparing two organisms because you can imagine, with the DNAyou have, it's all the time being mutated. It's being mutated by cosmic rays, by chemicals in theenvironment, actually by just UV light will, can cause mutations. And most of these mutations arebad, it's sort of like you need mutations to get evolution, to you know, advance the species, but it'ssort of like going to your car and whacking it somewhere with a hammer and, you know, there's achance you might improve it, but the odds are really quite small and probably you'll break it.

Hopes for the futureMary-Claire Kinghopes for cancer treatments

What all of us still hope is that the enormous amount of biology that we've learned, about thedevelopment of breast cancer and ovarian cancer, from knowing that mutations in BRCA1 andBRCA2 are involved, will eventually lead to treatments for those cancers themselves, and will in themedium term lead to ways of detecting very small breast cancers and very small ovarian cancers.That is, not just identifying a woman who is likely to eventually develop one, but actually detectingthe cancer itself, whether it's from a woman with an inherited mutation or otherwise. That kind ofwork is at least as big a challenge as the identification of the genes was originally, and many of us,both public sector and, I assume, private sector, are out to identify those sorts of tools, those sorts ofgene products on the surface of cells that we can see later on, those downstream consequences ofmutations in BRCA1 and BRCA2. It's an interesting time. None of what we do could be done withoutDNA science.

Finding cancer genesMary-Claire Kingsearching for candidate genes in families with breast cancer

For me, since I was looking at this from a genetic point of view, it was… The level of ignorance wasokay because what I was asking was, “Can I follow chromosomes?”, and the fact that the mechanismsof cancer were not understood wasn't critical to my being able to proceed. What I needed was agreater understanding of DNA itself. So what we were looking for was some fragment of DNAflagging some chromosome somewhere in the genome, that was shared by all of the cancer cases inthe family, all of the women with cancer, and was not held by branches of the family where there hadbeen few women with cancer. What made that sort of search possible in families like this, and therewere 22 of them in our original study, was the development of a genetic map. When we first startedthe work, we and lots of other groups like us were developing the map as we went along. By the timethat BRCA1 was ultimately identified in 1994, the map was essentially in place, thousands of markershad been placed on the map. What had taken me 17 years to do between the…from the earlyseventies until 1990 could be done now in weeks. The existence of the map was a phenomenal tool.So what we need to solve these problems are families like this who will talk to us, keep talking to us

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over the years, and let us work with their DNA, maps, genetic maps, and then the capacity to knowwhat sequence lies between markers in the genome.

Early cancer studiesMary-Claire Kingstudying cancer prior to understanding its mechanisms

When I began working on the problem of breast cancer in families, we knew very little about thecauses of cancer. There were elegant studies of the morphology of cancer cells and how they differedfrom normal cells, but this was before the discovery of oncogenes, it was before the discovery oftumor suppressor genes, of which, of course, BRCA1 and BRCA2 turned out to be examples. It wasbefore the discovery of the critical role of DNA repair and mutator genes. So the mechanism of how anormal cell becomes a cancer cell and how that happens in breast epithelium or ovarian epitheliumor in the lung or in the colon, was not well understood, although the phenomenon was beautifullydescribed.

Limitations of testingMary-Claire Kingcurrent status of testing for cancer genes

Where we stand is that it's possible to identify for some women in families with very large numbersof breast and ovarian cancers, whether they are at risk or not. That is to say that if a woman has aconventional sort of mutation in one of these genes, it can be found. And the great majority of suchmutations are found by commercial testing. That commercial testing is expensive, and it’s expensivebecause it is labor intensive to do. However, that sort of testing misses many mutations and it missesmutations not because it’s improperly done – I think it’s done very well – but because many of themutations in these genes are inherently impossible to pick up for biological reasons, using the kindsof technology that are being applied to them. We need a much more open field that allows andindeed encourages the development of new technologies for identifying these cryptic mutations inthese genes and in other genes, it’s not unique to BRCA1 and BRCA2. That’s the next challengewithin, in the very short term.

Patenting chaosMary-Claire Kingon knowing the function of a gene before you patent

The situation with gene patenting now in the[United] States at least, the one I know best, is chaotic.If I identify a bit of DNA, for whatever reason and if I search hard enough, on average that bit ofDNA has probably been patented three times over. The idea of patenting bits of DNA or even wholegenes whose function one doesn't know, is rampant out there. The NIH and the Patent Office aretrying to bring some order to this, so that the person who wishes to patent or the firm who wishes topatent, must at least have some sense of the function, the non-trivial function, of the gene that theywant to patent.

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Commercial patentsMary-Claire Kingon patenting genes for commercial purposes

I'm a great believer in the logic of having a patent on a new thing that allows one to make or dosomething differently, including medications. I do not put genes in that category, to me genes arenatural products, it's like patenting your left thumb. Do I think this view is going to obtain? No, ofcourse not, we've gone too far down the road of people being able to patent genes. But from the pointof view of a person investing their money in a firm, the existence of the gene is decades away from aproduct that will make a lot of money for that company. The goal of a pharmaceutical company or ofa biotech company that's going to partner with a pharmaceutical company, is I should think, to havea product that actually allows one to treat disease, or to detect it very early on. The gene is a veryearly step in that process. The consequence of that is that when companies are set up to find genes,they are almost forced to be less than completely clear to their investors about the amount of timethat it's going to take for even the most elegant result to be translated into something useful.

Humans & chimpsMary-Claire Kinghumans and chimps share around 99% of their DNA

So we used, we took bits of blood from our friends, our human friends, and bits of blood from ourchimpanzee friends and compared them. And, using the techniques of the time, it was absolutelyclear ,even to a person with very poor hands in the lab like me, that these samples were very, verysimilar. And we can do the calculations. It was straightforward to do and they were 98, 99% thesame. And we thought, how can this be? And we used several different ways of approaching thequestion, and it was always 98, 99% the same. But clearly, humans and chimpanzees are socially verydifferent and objectively we made measurements of other people in Alan’s lab, made measurementsof bone lengths of humans and bone lengths of chimpanzees. And at the level of those sorts ofanatomic differences, humans and chimpanzees are not 99% the same and we’re properly classifiedin quite different taxa on the basis of anatomical differences. On the basis of behavioral differences,similar as we are, we’re still appropriately classified in different families. But at the genetic level wewere very much the same. So we said what could this possibly mean, how can we reconcile theseeming discrepancy? And it seemed to us that the most logical way to reconcile the discrepancy wasthat there must be differences not in the primary sequences themselves in large number, but thatthere must be critical – rare but critical – differences in the DNA that lead to differences in thetiming of expression of genes during human development and during chimp development.

Studying DNA replicationArthur KornbergDNA polymerase: the key to understanding DNA replication

I work with bacteria because they're relatively simple compared to animal cells, they reproduce everytwenty minutes. We know much of their genetics and biochemistry. We still do not understand thenature of the more complex machine that includes the DNA polymerase, the enzyme that makes

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DNA, with many other parts that open the DNA, that prevent it from getting entangled, that makethese corrections when mistakes are made, rarely to be sure. And so how that whole structure, call ita replisome, something that has perhaps fifty working parts, we've not taken it alive yet, and itremains for the future research to understand this machine in its proper context and all its workingparts. And without that we really will not fully understand the health of a cell, the disease of a cell, ina very fundamental way.

Enzymes: DNA polymeraseArthur Kornbergthe catalytic capacity of DNA polymerase

You depend in every chemical reaction in nature, even the hydration, adding water to carbondioxide, on an enzyme to catalyze that reaction thousands of fold faster than it ordinarily would, andto direct it. And so we already knew that enzymes had to exist to catalyze and direct the assembly of adouble helix. And so DNA polymerase, rather unconventional in that it was not programmed to doone specific thing, but rather to take directions from a pre-existing DNA, but I would say overallminor compared to its catalytic capacity to match As with Ts and Gs with Cs, and to do it at anastonishingly rapid rate, thousands of times a minute with very rare mistakes.

DNA synthesisArthur Kornbergthe papers that first described the isolation and synthesis of DNA molecules

We submitted two papers in 1958, to the Journal of Biological Chemistry, which I'll say perhapsimmodestly, were classic papers, because we described each of the stages in which one made andisolated a deoxyribonucleoside triphosphate, the A, T, G, and C, described how the enzyme wasisolated, not really isolated, partially purified, but sufficiently separated from other components sothat there was no degradation of DNA, the need for DNA, the need for each of the four buildingblocks, without which virtually no reaction occurred, and the demonstration of the net synthesis ofDNA. And so those were described in great detail in these two papers, with the title “DNA Synthesis.”

Outcome of the HGPEric Landera new paradigm for studying biology

The Human Genome Project changes things so there are no more genes you don't know about, orparts you don't know about, and we haven't fully digested those implications, we don't quite have allthe last little bits filled in, but two generations from now our scientific descendants, that is ourstudents' students, will look at biology the way chemists look at the periodic table, it's there in thefront of the book and there's no more elements. And the Human Genome Project, it'll just be there inthe front of the book.

Junk DNA & evolutionEric Landerjunk DNA may have important evolutionary functions

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All of these interruptions of the genes probably play key roles in physiology, in regulating the gene inimportant ways. They allow the organism in one tissue to splice up a message one way, and in othertissues to splice up a message another way. And then on a grander scale, they allow evolution totinker more easily. You can have chromosomal breaks that make a new combination of chromosomalsegments and most of the time they'll occur in this useless junk stuff in between and therefore amessage made there will still be spliced. So it allows random breaks and reconnections to give rise tofunctional genes because of all this spacer stuff. So it looks stunningly inefficient, it makes your headhurt to think of making two million letters of DNA and throwing it almost all away. But evolution isin this for the long term and it really thinks about how these mechanisms can be used over longevolutionary periods, different tissues etc. It’s… I mean, again, it tells us that we are very far frombeing able to design organisms and that we need to sit at the feet of evolution for an awful lot longerand learn what it's learned over three and a half billion years before we have the hubris to imaginethat we really understand life.

Smell receptorsEric Landerevery gene has a distinctive evolutionary history

Every gene has its own distinctive history. You take the smell receptor genes, we have a thousandsmell receptor genes in our genome, and that says that our vertebrate forebears were really, reallyinterested in olfaction as a sense. And then you look at them and you find out that two thirds of themare broken, they've all accumulated mutations in the past tens of millions of years, saying thatsomehow our more recent hominid ancestors lost interest in smell.

Bacterial vs. human genomeEric Landerhuman genes are organized in patches of information

So in bacteria, genes are really simple. They start, they run continuously, and they stop. It's reallyeasy, you get a long stretch of, oh I don't know, a thousand, two thousand DNA letters that arecontinuously coding for protein. The human genome, it's nothing like that. A typical gene will bebroken up into eight, ten, twelve, sometimes seventy little patches of information, each patch ofinformation might encode, oh I don't know, fifty building blocks, fifty amino acid building blocks forthe protein or so. That's a typical sort of size. And the problem is that they're spaced very far fromeach other. There's lots of stuff in between, some of it's junk, some of it is regulatory sequences.

Processing mRNAEric LandermRNA editing by the spliceosome

When the cell wants to copy a gene, RNA polymerase, the enzyme that sits down and copies a gene,goes to the beginning of a gene, the promoter, and it starts copying faithfully out the whole RNAcorresponding to the whole DNA, and maybe it might run fifty thousand letters, makes an RNA thatthen goes off into the cytoplasm. These fifty thousand letters actually only have maybe fifteenhundred letters of information in it, and so this editor comes along, basically I mean the genome is

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kind of a flabby text and it needs to be edited, and so this editor called the spliceosome comes along,grabs this very long message, of which there's not too much that matters, and starts at one end there,and figures out, by ways we really don't fully understand, where's the first cut it should make. Weknow a little bit about the sequence that's there, that it kind of recognizes, but we're not so good atperfectly picking those out. Then it holds that and it looks down the message for where it shouldsplice that to, and it sees some signal there and we know a little bit about what it's looking for… grabsthose two, brings them together, cuts, closes them up and then throws away everything in between.Then it's got to do that to the next bit, the next bit, the next bit, the next bit, till it gets down to theend.

Sterilization as a welfare reformPaul Lombardosterilization in the USA: used as a welfare reform

Among people who call themselves Eugenicists sterilization was thought to be the last effort to doaway with people who would generate social costs, people who raised the tax burden, people wholived in institutions for the dependent or for, as they said, the defective or the delinquent.Sterilization was meant to be a kind of welfare reform.

The Buck vs. Bell casePaul Lombardosterilization in the USA: Buck vs. Bell

In the history of American law, there is no case like Buck vs. Bell. It's the only occasion in which theSupreme Court of the United States has allowed surgery on a person who didn't wish to be operatedon and who didn't need the procedure. The case was built on false facts, false theories and falsescience and it was used as a model not only in the United States but all over the rest of the world.Following the Buck trial the precedent had been set for people to be coercively sterilized by the state.Unfortunately the provisions that were written into the Virginia law and lots of other laws like it werenot followed in the future. Many people were sterilized without having been told that that was theoperation they would endure, many others never learned that they were sterilized until years laterwhen they tried to have children.

Complex behaviorHubert Markl the honeybee as a model for complex behavior

I think the honey bee which they are now are going to sequence too, is in one sense a wonderful,simple and complicated organism because it's simple enough, as Randolph Menzel and others haveshown, that you can really see how genetics can program very complicated behavior. A bee's dance isvery complicated but it is almost completely genetically determined. But then environmentalinformation is added to it. There you have a wonderful model which is comparable to bird song orbird migration. And this isa very important message, that very complex behavior need not beinfluenced by a large complexity of genetic systems. It can be few genes, switched on in the right kindof collection, and in the system they can have a lot of impact. And there we come closer to what I feel

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will turn out to be true in the human beings, that…and the comparison with the monkeys and thechimps will show us even more of that. It's not a new set, entirely new set of genes, it's the way howthe interplay of the genes builds a different system, which makes really the difference. And there, Imust say, I am looking forward as a scientist to a very glorious future.Genetic pre-wiringHubert Marklbehavior can be both genetically pre-wired and learnt

An insect is fully wired, so to say, is born fully wired, and then if, if the legs somehow are failing todevelop right, then they stumble, whereas mammals are exceedingly adaptable in that. Now on theotherhand, if you look at bird flight for instance,we know exactly from [G. S.] Spalding's time thatyou can bring up a bird under so tight conditions like a swallow, that it has moved a wing, andwhenthe time has come it has everything wired in. So we have both conditions, like in bird song,which you know also so well. It can be completely pre-wired, it can be completely learned, and itseems to me that as far as the human being is concerned, we have…well, maybe even exaggerated,but when did all the way to use the full flexibility of a brain which is using as little pre-wiring aspossible, and then dropping the routines you don't need.This is true for the cognitive part,In theemotional part we have a lot more of wiring, when you talk about hunger, when you talk about sex,when you talk about aggression, you talk about sleep, then you see in the limbic system and in themesencephalic systems, there is a lot which is there which is very similar, maybe even to reptiles orat least primitive mammals.And we have to realize that the emotional things are very stronglygenetically pre-wired because nature doesn't run the risk that you have the wrong emotions.

Classification and valueHubert Marklconfounding genetic classification with human worth

You begin by accepting that there is something scientifically sensible about classifying peopleaccording to their contribution from different genetic heritage. If you, this is a first step, and you endup very easily then in classifying people according to their worth and I think we have to make it veryclear. Of course there are population differences, of course black people look different from whitepeople, and we are a deficiency mutation and others look like Mongols do, and so on. But this hasnothing to do with the value of human people. And these, to confound these things is a mosthorrifying thing and apparently if you are in the right social surrounding, where that becomes notonly acceptable but even the command of the time, then you end up with doing terrible things.

A better understandingHubert Marklimproving our species with better education

What we can do to improve human beings, does it have to do with what we understand from thegenome project or genetics, yes, but in a different way, not by implanting better genes or so, it will bea better understanding what you can do for a given genetic condition which you have inherited, inorder to fulfill its potential. We probably will learn much about the conditions of learning, learning isthe most wonderful thing in the world and we think, well this just happens, but I think we can do a

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lot to make it happen better. So we do have a way to improve the human being, we call it education.

DNA is the genetic materialMaclyn McCartythe experiment that identified DNA as the genetic material

You realized that you were, the substance that was in these extracts not only caused the change but itapparently was also reduplicated, because you took, could take transformed cells and extract thesame thing from them that would transform other cells. So you had something that was, caused achange, and the stuff that caused the change was also reduplicated. This, if you think about it, has avery genetic kind of thinking… since it was known that DNA was in the nucleus in the cell andcertainly was in the chromosome, so that the unit that carries all the genetic material so that it madesense to think about DNA.

Living with sickle cellKatreece McGheehow sickle cell has affected her life

Katreece suffers from a disease called sickle cell anemia that affects her hemoglobin. Hello Katreece.How are you? It sometimes causes her a great deal of pain. What kind of pain are you having? Chestpains Chest pains. And on a pain scale from zero to ten what would you say that was? About a nine. Anine. The pain is worse than breaking a bone or fracturing a bone, it’s a real extreme kind of pain.The only thing that comes to mind is somebody stabbing me. Have you been coughing a while? No.Have you had any trouble breathing at all? No.I was always stopped from doing what other childrendid. I couldn’t play in the snow all the time, I couldn’t go swimming on certain days. It really took atoll on me at once because I always wanted to do it and I’m like, well, why me? What happened here?

Inheriting sickle cellKatreece McGheehow she inherited sickle cell

The disease gets its name from what happens to Katreece’s red blood cells when she experiences anattack. They become sickle shaped and these distorted cells get stuck in small blood vessels so thatparts of her body don’t get the oxygen they need. Sickle cell anemia is an inherited disease.Katreece’s parents each have one copy of the gene for normal hemoglobin and one for the sickle cellkind. They get by fine. But unfortunately Katreece inherited the faulty copy from both her parents, soshe has the disease. For now, all doctors can do is help relieve the pain. But there is hope thattechniques being developed today will lead to therapeutic interventions for genetic diseases such assickle cell anemia.

Replication modelsMatthew Meselsonthe different models proposed for DNA replication

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Well we tried to find out if DNA replicates semi-conservatively, or dispersively, orconservatively.And that means, do the two strands separate but remain as single strands indefinitely,and each become associated with a newly-synthesized strand at each generation? Or do they comeapart and get dispersed, or do they stay together, the two strands indefinitely, making only brandnew DNA molecules alongside themselves?

The correct modelMatthew MeselsonMeselson and Franklin Stahl's experiment to determine the correct DNA replication model

So we start the experiment by growing the bacteria for a long time, many hours, many generations,in heavy nitrogen medium, so everything's heavy nitrogen. And now we put a little bit of that bacteriainto some Dupenol detergent to liberate the DNA, we put that in the ultracentrifuge and wecentrifuge it and all the DNA is heavy, it forms a single band. Meanwhile most of the bacteria havebeen left behind in the culture tube, and we centrifuge them down and resuspend them in lightnitrogen medium. So now any more DNA that's going to be made is light, not heavy, and we see that,as time goes on, there builds up a new band until at one generation, one full generation after thetransfer from heavy to light, all of the DNA is at this new density, which is the density we expect forDNA with one heavy strand and one light strand, namely hybrid DNA. And then as time continues togo on, back in the culture tube where the bacteria are growing in light nitrogen medium, still anotherband begins to appear, that's fully light DNA. Both strands are nitrogen 14, the light kind of nitrogen.And now as time goes on, the heavy strands that are inherited from way back there are neverdestroyed, they're in a sense immortal, but as more and more DNA is made it can be made only outof light DNA, and so the light-light DNA, two strands both light, begins to become more and morepredominant in each sample, until…we ran this experiment in this case to four generations. So that'sit.

Can genetics provide answers?Benno Müller-Hillgenetics may not provide the answers we seek

I am now going to say something which is outrageous and which presumably Jim would not follow.And this is, I think that until now very, very little has been done in this type of human genetics,schizophrenia, manic depression, always some loci have been identified and then the locidisappeared and there is nothing left. Maybe there is something, there is now some new loci, but Iwould, I am not quite sure whether ever anything will be found, whether our brain is not simply toocomplicated, that genetic determination does not exist, we become crazy or not crazy or whatever,on, not via simple genetics. And I think what, if you really would like to know, either what theunborn is you have to be able from a DNA analysis to predict how it will behave. And I think we aremiles away.

Who should decide?Benno Müller-Hillmaking life choices and economic considerations

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The parental choice without any, not being enforced by economics, yeah, because there is no otherone, there are two possibilities, either you say you don't do anything, yeah, every, putting the fingerin this development, no, absolutely no. So if you say you allow it, then there are two possibilities,either you leave it, two extreme possibilities, either you leave it completely to the individual which isinvolved, which is the mother, or you leave it to the medical doctors and to the State. And I would notlike to leave it to the medical doctors and to the State, so I would leave it to the individual. Butwithout economic pressure, the economic pressure comes, then things are totally different, yeah.

Polymerase chain reactionKary Mullisdiscovery of the PCR technique + ANIMATION

This procedure will copy the DNA that is between the place where these two little short pieces of itare, they're stuck to it, ‘cause their sequence is right to do that. And if you run this process once you'llget an extra copy of the thing that you were trying to figure out what it was in the first place, so youhave twice as much. And if you do that again, there's nothing that stops me from doing it again, Irealized, that was the, and I said my god, if I did that again I'd have four times as much, and if I did itagain I'd have eight times as much, and I could keep doing that till thirty times would give me abillion times as much of this particular little sequence that contains the information that I'minterested in, to find out whether or not this fetus is going to have this particulardisease...Polymerase chain reaction, or PCR, uses repeated cycles of heating and cooling to makemany copies of a specific region of DNA. First, the temperature is raised to near boiling, causing thedouble-stranded DNA to separate, or denature, into single strands. When the temperature isdecreased, short DNA sequences known as primers bind, or anneal, to complementary matches onthe target DNA sequence. The primers bracket the target sequence to be copied. At a slightly highertemperature, the enzyme Taq polymerase (shown here in blue) binds to the primed sequences andadds nucleotides to extend the second strand. This completes the first cycle. In subsequent cycles,the process of denaturing, annealing, and extending are repeated to make additional DNA copies.After three cycles, the target sequence defined by the primers begins to accumulate. After 30 cycles,as many as a billion copies of the target sequence are produced from a single starting molecule.

Public & privateGene Myerscomparing methods used by the public and private teams

In the public project the approach was to basically divide the genome into about 25,000 pieces, largepieces, and to first figure out and map each of those pieces to the human genome. Each of thoseindividual 25,000 pieces then, was then shotgun sequenced, so they used shot gunning, they justused it at a scale of about 150,000 letters. The problems would typically involve about 2000 pieces,whereas we side-stepped the idea of actually mapping these 25,000 pieces and instead said, let's justdirectly shotgun sequence the human genome, so let's blast it into pieces, grab 500 letter pieces andlet's go ahead and solve that single assembly problem. So we had a single shotgun assembly probleminvolving 40 million pieces, so the basic difference is that in the public approach they solved 25,000problems of size 2000. We solved one problem of size 40 million.

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An overviewGene Myersusing computers to assemble genomes and interpret data

There are two kinds I think of bioinformatics work, one is what I would call purely in silico work,where the idea is that you basically are taking the data that's available on the computers and trying tomake inferences about genes, about the functioning of genes, about the signals that are present. Theother kind of computational activity in bioinformatics is about interpreting experiments, and forexample we have two very large examples of that, we're doing all of the computational work forinterpreting what comes out of our proteomics factory, and I consider the whole genome shotgunsequencing and the subsequent assembly step, to be another example of that type of bioinformatics. Imean if you think about it, there was a large experiment, we probed the human genome forty milliontimes and then there was a huge computational problem, a kind of a, the king-sized problem.

3 DNA bases = 1 amino acidMarshall Nirenbergthe mathematician George Gamow's idea that three DNA bases encode one amino acid

He told me that he went down to his driveway to the mailbox to pick up the mail, and picked up thatissue of Nature that contained Watson and Crick's article on the helical nature of DNA. And he readit while he was standing at the mailbox with one arm on the mailbox, and immediately thought thatthree bases in DNA corresponded to one amino acid. There are four kinds of bases in DNA, twentykinds of amino acids in protein. And so, taking them three at a time there are64 possiblecombinations of the three bases.

The role of the ribosomeMarshall Nirenbergribosomes recognize a triplet code

One time I asked myself the question, what's the smallest word that would be functional, that wouldbe recognized? And I thought that maybe a triplet, three bases alone, might be recognized onribosomes by the appropriate species of transfer RNA with, the appropriate amino acid attached toit. And we, I tried this and the very first experiment worked, and so we used a very simple, a differentkind of assay, a very simple assay to determine the base sequences of codons by measuring thebinding of radioactive amino acid attached to transfer RNA, to recognized on, bound to ribosomesand recognized a triplet, three bases.And we could separate the complex from the unboundaminoacyl tRNA by adsorption on filters.

Cracking the first codonMarshall Nirenbergdeciphering the first amino acid codon

We were looking for a synthetic polynucleotide, an RNA that contained only one kind of base insteadof the four bases. We were looking to see if it would stimulate the synthesis of protein that contained

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only one kind of amino acid. That's what we found. So we found that a series of Us in RNAcorresponded to… it was the code word for phenylalanine in protein. We deciphered the first, I meanwe'd broken the code, we'd deciphered the first codon, the first word that in nucleic acid languagethat corresponds to an amino acid in protein. It was fantastically exciting and it opened the wholedoor to deciphering the rest of the code.

Universal codeMarshall Nirenbergall forms of life use the same genetic instructions

This finding that the code is universal had a terrific philosophic impact on me. I mean I kneweverything about evolution at the time but these findings were so immediate and so profound that itjust had a great effect on me philosophically, because I understood that most or all forms of life onthis planet use the same genetic instructions, and so we're all related, we're related to all livingthings. And when I came in the garden and saw the plants and the squirrel and some of the birds, itreally had a profound effect on me, which lasts to this day. I think that this feeling of being one withnature is very real, and in fact it's very true, we all use the same genetic language.

Cracking the codeMarshall Nirenbergdeciphering every triplet code

Gobind Khorana is…was, at that time, one of the world's best organic chemists working in the fieldof nucleic acids, highly experienced and superbly equipped to do this. He synthesized the 64 tripletschemically and he also synthesized repeating polymers with known doublets, repeating doublets ortriplets as well, and towards the end he used them all to determine nucleotide sequences also. *Sobetween the two labs, he used the ribosome binding assay that we had established, and between thetwo labs we determined all of the nucleotide sequences of RNA codons.

Protein synthesisMarshall Nirenbergsynthetic RNA stimulates protein synthesis

Everything fell into place, everything worked. I mean we soon found that, that adding RNA whichwould have been prepared from ribosomes, to cell-free extracts of E.coli stimulated amino acidincorporation into protein. All the controls were there, even though the stimulation was tiny. It waslike, like thirty counts per minute above a background of maybe a hundred or eighty counts perminute. It was a small stimulation but it was real, and when I saw that, I mean the very firstexperiment, I knew it was true. I knew that template RNA was stimulating cell-free protein synthesis.I jumped for joy because we had really found that RNA, messenger RNA could stimulate proteinsynthesis.

Neandertal & human ancestrySvante Pääbohuman origins and our common ancestry with Neandertals

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What we think we know about the origins of modern humans today is that we go back to a rathersmall population that lived sometime around 100 or 200,000 years ago in Africa where somechanges happened that we don’t quite know what they are actually, that made this group expand andreplace other forms of humans elsewhere, perhaps interbred a little bit with those groups but not togreat extents and when we trace our ancestor, common ancestor with a Neandertal back, that goesback much further about two or three times further back. So around half a million years ago, weseem to have shared an ancestor with the Neandertals.

Neandertal DNASvante Pääbocomparing Neandertal and modern human mitochondrial DNA

So what we did was to compare the sequence that we were able to retrieve – short, short pieces thatwe puzzled together from this Neandertal – and compared it to humans worldwide. And what wefound was that it was not at all more similar to people in Europe today than to people livinganywhere else. In fact this sequence sort of branched off before the common ancestor of all themitochondrial sequences of people living today, no matter if they live in Europe, in Asia or in Africa.So it seems that, or we know in other words now, that Neandertals did not contribute anymitochondrial DNA to people living today.

ProteomicsScott Pattersonstudying proteins to understand disease

So genes make proteins, and proteomics is the study of proteins. What we want to do is identifythose proteins that are expressed in disease tissue that aren't expressed in the normal tissue.Utilizing all of these efforts of cell biology, protein chemistry and then in the mass spectrometry labhere, we can identify those proteins that are over-expressed in disease tissue such as lung cancertissue, compared to the normal lung cells. We need to identify those, and to identify those proteinswe match back to the genome sequence that we'd already generated at Celera, and leverage thatinformation. Of course it's not just the wet labs that you're seeing here that are part of the proteomicseffort, we also have significant computational resources at our disposal to match all of this data intothe genome and to gather all of the other information that's available in biology till we fullyunderstand these potential targets for therapeutic intervention.

Differences & similaritiesRobert PlominDNA variations result in differences between individuals

It's really amazing to think that the basic mechanism is relatively, you know, simple, you know, it'snot to take away from Jim Watson's famous discovery of DNA, but it's quite amazing that there's thistriplet code that codes for twenty amino acids and they produce all the proteins that we are and inour bodies and in our minds as well. Neurotransmitters are coded directly from DNA. And we knowhow that mechanism works, and we know that some of those DNA bases change, they mutate for

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people, that's where all this genetic variation comes up. It's, you know, it's a relatively small portionof the DNA. You and I are the same at 99.9 percent of our three and a half billion nucleotide bases ofDNA. But it's that point one percent, one in a thousand DNA bases that make us different, and thoseare the ones we're after because we're interested in things that make people different, why are somechildren reading disabled, for example.

Heritability of behaviorsRobert Plominall behavioral traits have a heritable component

So twin studies took autism from thinking about it as an environmental disorder as to thinking aboutit as one of the most heritable disorders around, and now people have begun to identify some of thegenes that are involved. So that's one example of a trait that wasn't thought to be heritable, and onthe basis of twin studies it's now realized that it's highly heritable. A counter-example, though is, forexample, alcoholism, which has been thought to be almost entirely genetic, partly because it wasowned by the medical establishment and people assume, well, it's biological therefore it must begenetic. But the twin studies, the large ones, good ones, have only been done in the last decade andthey suggest that, although there is some genetic influence, it's not terrificallystrong for alcoholism,especially for females. Now there's lots of examples like this. The point I'm trying to make is that youcan't assume what is heritable or isn't heritable.I mean ulcers, people say “Oh well that's stress,environment.” Well, it turns out there's a strong genetic component to ulcers because some peopleare genetically more susceptible to that stress and it comes out in ulcers. So you can't assume what'sheritable. You have to assess it, and the twin method is a good way of getting at that. But the mostimportant point from all of this research, is that genetics is important just about everywhere we look.There's no behavioral trait that has been reliably shown to be zero heritable.

Nature vs. nurtureRobert Plominhow much of our behavior can be attributed to genes?

Well, how much is nature, how much is nurture? The answer is, nature's important wherever welook, genetic influences are important. But for complex traits like behavior, like mental illnesses orcognitive abilities or disabilities, it’s not all genetic, and onaverage you could say something like fiftypercent of the differences among people, half of the reasons for, say, reading disability or languageimpairment,is due to genetic factors. So it's important to emphasize, genetics is very important, but,the second message is, genetics isn't the whole story, there's a lot of environmental influence and it'sabout 50-50 on average across a lot of these abilities and disabilities.

Designer babiesRobert Plominintervening in a child's future at a genetic or social level

Many people are concerned with designer babies, for example, and if you find genes for these traitsparents are going to use them to select babies, say prenatally for example. And I think with anyimportant knowledge there are also important problems that knowledge can create, and we have to

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discuss those, I hope rationally and try to prevent the problems and maximize the benefits from thissort of work. I think though, in terms of genetic engineering, these complex traits like autism,schizophrenia, reading disability, language impairment, the evidence suggests that they're influencedby many, many genes, so it's going to be hard to get levels of prediction at an individual level, that is,you could make a prediction in society, you know, on average, children with these genes are morelikely to be reading-disabled than the other children, and you might develop programs to try andintervene at that level. But that's quite a different level of policy than trying to predict this one child'spotential for becoming reading disabled or schizophrenic or autistic or whatever.

Prediction & preventionRobert Plominpredicting and preventing behavioral problems

My main interest in doing this sort of research now is to find the most heritable components andconstellations of these disorders. You can't just sit in an armchair and theorize about what's relatedto what genetically and what ought to be heritable or not be heritable, you have to do this sort ofresearch. And then once we find the most heritable aspects, say of language impairment, my maininterest is to harvest the fruits of the human genome project and begin to identify some of thespecific genes that are involved. And the reason why I want to do that is because I think it will allowus to understand some of the basic processes that are involved, perhaps in the brain, but I think evenmore quickly it will help us begin to predict children who are likely to have problems and perhaps todevelop interventions that allow us to prevent the problems before they occur. That is, for readingdisability, rather than waiting until children go to school and they fail repeatedly, and then you say,“Oops, reading disability” perhaps you could predict that with some genetic markers earlier. You'vegot to avoid problems of labeling the children and all of that, but it would give you a shot then atpreventing problems before they occur, and surely preventative medicine has to be the way to go.

Twin studiesRobert Plominthe genetic basis of cognitive traits

Well, the twin method compares identical twins who are like clones of one another, so they'reidentical genetically. The other type of twin, fraternal twins, are fifty percent similar genetically, likeany brother and sister. So the twin method compares these clones, identical twins to fraternal twins,and if genetics is important you'd expect the identical twins to be more similar, say for languagedevelopment, than the fraternal twins. And that was the case, for language abilities of many differentsorts, identical twins are more similar than fraternal twins, suggesting genetic influence. But what'snew in the last few years is to look at language impairment, the children who are quite slow indeveloping language. And ifanything, those results show even more genetic influence, that isidentical twins areeven more similar and fraternal twins are even less similar for these sorts ofcognitive disabilities.

Basis of complex disordersRobert Plominunderstanding the genetic basis of complex traits

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I think the most important thing in all of this is that DNA is important, not just for bones andbiology, but also for behavior. We're at a point, thanks to the genome project of beginning to identifysome of the specific genes that are involved in these complex traits. And that I'm sure is going to leadto some very major advances in understanding the origins and the treatment of some of the mostimportant disorders in our society.

Unused toolsRobert Pollackgene technology and its possible uses

On one hand you have in principle the capacity to use this technology to create organisms thatpenetrate all immune and pharmacological defenses and kill people at will. On the other hand youhave, through this technology, the capacity to enhance the immune system and createpharmacologicals that interfere with malaria, which infects one person in four today, or TB[tuberculosis] which infects one person in three today on this planet. Where do we put our money,where do we put our work, where do we put our public awareness of what's possible. We put it to thecertainty that rogue countries are doing the former, and check the budget for what we're doing on thelatter. But no, the technology is neutral, the technology is safe and neutral, the uses of it and theconsequences in subsequent 25 years have been appallingly stupid, relative to the risks. These aregreat tools that lie in the toolbox, untouched, as far as I can see.

Risks of DNA recombinationRobert Pollackpotential risks associated with recombining DNA

I had in my class a student who was a graduate student of Paul Berg's in California. And she wasthere to learn how to do the transformation assay, so-called, ones in which one adds SV-40 virus tocells in a dish and watches them heap up and become released from the growth controls that calmnormal cells down. And she was very good, very bright woman, and proposed to us that we consideras a class the significance and utility of work she was doing to move the genome of SV-40 byrecombination using restriction enzymes, into the E. Coli bacterium, which was not the medium ofmy interest, but rather the bacterium which lives commonly in one's gut and is the major bacterialtool for studying genetics and DNA modifications, then and now. I posed to her the simple question,whether she had thought about the fact that she was bridging evolutionary barriers that had existedsince the last common ancestors of bacteria and people, by putting a, a viral genome from a primatevirus into a bacterium, and whether that might jump a species barrier and cause someone to developcolon tumors from transformation of their colon lining as this bacterium established itself in theirgut.

Gleevec: first trialsBud and Yvonne Rominethe first patient in the Gleevec trials

Now this is the very first day, fact is it's June of ’98, I don’t have the exact day, it’s when the clinical

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trials started and Bud was the first patient to take this and it was 25 milligrams. And there is apicture that was taken of him and Dr. Druker wishing him luck. So we’re pretty proud of that picture.And then this guy over here, he’s showing you what the pill looks like, so at that time it was a numberbut now it's known at Gleevec. Yeah STI-571.This is a chart of blood, white blood counts that wastaken. This is about a year before he started – at that time it was known as STI-571. Yes, that’s it. Andhe was in the 35,50 thousand range. 60 thousand… white blood count. And the white blood countwhich is normally, I think it's about ten thousand is about the highest a person would get on it. Thenhe started taking this STI-571 which is Gleevec and about, about 17 days he was back down into hisnormal range on his white blood count and he’s been normal ever since. And this is just, what animportant thing, what a breakthrough! Some things make you sick, you know, I mean, but this hereeven most everybody can take it, even small children. And it doesn’t make you sick, which iswonderful so it's wonderful stuff. Yes sir, good old Gleevec.

DNA variationMark Skolnickmeasuring DNA variation: techniques and applications

When people refer to restriction fragment length polymorphisms, also known as RFLPs, or SNPs,single nucleotide polymorphisms, these are just different techniques for measuring DNA variation,which all of us have in the very long three billion base DNA sequence. So there are the DNA basesthat make up the genetic code, the genes that cause the proteins, and variations both within thegenes and between the genes, can be used to follow patterns in families. Another use of them thatmany people are aware of, is for DNA fingerprinting, identifying within a blood sample a specificpattern of variation which is characteristic of an individual. A very well-known example of this wasthe O. J. Simpson trial, where the DNA fingerprinting, I think, or RFLP technology, was discussed alot in the popular press.

Identifying BRCA1Mark Skolnickfinding and cloning the first breast cancer gene: BRCA1

So the process of finding a gene is very complicated. You get a signal that you have found somethingdifferent in a gene in a family, and the first signal that we got for BRCA1 was actually a rather weaksignal. It caused a slightly different protein to be made, but we couldn't be convinced that thisprotein actually was an abnormal protein that would cause a disease. So there was excitement, therewas jumping up and down, but we knew we needed to find in another family a stronger changebefore we were really sure that we had the gene. Because, in this process, there are what we call genescares, where you're looking at a gene, you see some changes, you hope that those changes mean thatyou've found the abnormal gene, but in fact as time goes on you discover that they're just normalvariation, normal DNA variation in normal individuals. So it took some time for us to really be surethat we had the breast cancer gene and every time we got more evidence that this was it, there wasanother thrill, more excitement, and then once we were sure that we had the gene, then another racewas on, that was the race to find the whole gene, because the prize was the whole gene.

Looking for BRCA2

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Mark Skolnickfinding the second breast cancer gene: BRCA2

Well, just after we found BRCA1, we were then able to separate the whole set of families we had intothose that were linked to BRCA1 and those that showed no linkage. So one member of our team,David Goldgar, teamed up with another player who hadn't been much of a player in the BRCA1 race,Mike Stratton, in an attempt to actually look at linkage information for those unlinked families, forthe families that were not likely to be BRCA1 gene-carriers, and to look for the next gene, BRCA2.And why did we do it? We did it because it was the next logical step. Your families are now purifiedin that you have a clean set of just BRCA2 families. If we're going to offer a diagnostic we want tooffer a complete diagnostic, that is for both genes, not just one gene, so it was the logical next step forus.

Counting DNA mutationsMark Stonekingwhy the number of mutations in mitochondrial DNA is an underestimate

And so if you have a particular nucleotide at one position, say an A, and when it mutates it willchange to a G, a C or a T to something else. Now if you look over a long enough evolutionary periodof time, that particular point in the DNA segment has a probability of mutating again and so, andbecause there’s only three choices that it can mutate to and be seen as a difference there’s a highchance that it will mutate right back to what it was before. So you could have an A mutate to a G,then the G mutates back to an A and so you’ve had two mutations, and yet if you only compare, if allyou see is an A in one species and an A in the other you will say there have been no mutations. Sowhat we actually observe, the number of differences that we count is an underestimate of the numberof mutations that have actually occurred.

The divergence of NeandertalsChris Stringerfossil evidence shows that Neandertals diverged from modern humans

Well my work, and the work of plenty of other people as well, suggests that the Neandertals areclosely related to us, but they may well be a distinct species. They diverge from our line of evolution.On the fossil evidence, sometime in the middle Pleistocene, we might say perhaps three hundredthousand to six or seven hundred thousand years ago, their line of evolution diverged from ours. Andthe interesting thing is that the mitochondrial DNA that’s been recovered from only a handful ofNeandertal fossils is consistent with that view. So this mitochondrial DNA — even though there’sonly small bits of it — it suggests that first of all the Neandertals had their own population variationwhich was comparable with the variation we have today, but it was distinct from our variation. TheNeandertals are no closer to Europeans, for example, today, than they are to say Australians orAfricans of today. So they are a distinct lineage. They started to go their own way in the middlePleistocene, and the DNA would suggest perhaps something like five hundred thousand years ago,they started to evolve genetically in a distinct way from modern humans.

Reading our own code

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John Sulstonreflecting on our evolution

I mean one thing I do like is to reflect on the fact that it took four billion years for life to emerge andevolve to the point where one particular living organism was able to read its own code ofinstructions. And that I think is a most wonderful philosophical point. There'll be a much morewonderful philosophical point when we understand how the code of instructions works, but at leastwe've read them out on the tablets, you see, at this point.

A free flow of informationJohn Sulstonmaking sequence public to pre-empt the patent

Remember that we did have to respond to the announcement from Celera [Genomics] in thebeginning that they were going to patent some fraction of the human genome. I hope that nobody'sdenying that, because that is very clearly on the record that that was going to happen. We thereforeknew that if we wanted to head off this and make the genome really free for all of humanity, we weregoing to have to make sure that there was prior art out there, that was the only way of doing it. Imean, we can't, western, unfortunately although I disagree with it in many respects, western patentlaw states that if you get in there and you file a patent and so on, you've got it, and it's going to beextremely difficult to break that. And if he, if they were able to do that on substantial parts of thehuman genome and it would be in their hands how much, then we would all be in very bad shape.And so we felt that the only way to deal with this was to make sure that the public domainsequencing went ahead as strongly as necessary to preempt the gene patenting, to maintain a flow ofsequence into the public domain, so that people who needed it would have full access to the humangenome.

Evolutionary relationshipsJohn Sulstonthe conservation of life processes

Well the key thing to have in mind is the unity of life. If every life form were specially created as somebelieve, and they have no particular relationship to each other, then of course there wouldn't reallybe much value in studying more than the one you're actually interested in. But the truth is, that life isvery, very unified and it's unified, all the indications are, because of the evolutionary process, that itall goes back to a common ancestor four billion years ago. And it turns out that an awful lot of lifeprocesses have been conserved, so that you can look in bacteria and find that half the genes haveclear counterparts in a human being. You can look in the nematode. Of its twenty thousand genes,half of them have clear counterparts in the human being, and all the way up it's the same. So Natureis not reinventing, it's actually reusing, and it does more than that, it reuses whole pieces ofmechanism.

Human genome patentsJohn Sulstonthe human genome sequence is not a basis for a patent

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We also established the principle similarly to preempt silly competition that people would not betaking patent protection, IP [intellectual property] protection, on this sequence. That in itself, I meanthis is another strand if you like, the whole business of intellectual property in the human genome,but I think it was clearly understood by people in the room that this is not something that should bepatented, you know, because it's too basic, we don't know that much about it, it's not really the basisfor a patent application.

100 km of DNAJohn Sulstonthe length of the human genome

DNA is very long, it's a seriously long molecule, it’s … One copy of the human genome is one meterphysically in length. But one nice thing is, if you imagine the thickness of the DNA in the humangenome scaled up to the thickness of a piece of cotton, then it would actually be one hundredkilometers long, a single copy of the human genome.

After the HGPJ. Craig Ventera new foundation for science

Graduate students that are starting science today don't even have to think about ever cloning a geneor purifying a protein, they just instantly download that information. You know, I think what could Ihave done with that extra decade of my career, had I been able to just download that informationinstead of spending ten years trying to get it. It's a fundamental different starting point, it's a newfoundation for all of science.

Solving a problemJ. Craig Venterdeveloping the tools to sequence the genome

It's even more amazing that all of it worked and all of it came together, when we now see clearly withhindsight all the things that could have failed. When we first started Celera the sequencing machinewas an engineering prototype, we actually never saw it work. The mathematics, half a million lines ofcomputer code all were written prior to the sequencing of the genome. They didn't exist before, themathematics didn't exist before, they were all created in real time.

A recent common ancestorDouglas Wallacemitochondrial DNA confirms a recent common ancestor for modern humans

So the key observations in the mitochondrial DNA that were most surprising at the beginning wasthe realization that all human females, and therefore all of humanity is related through one bigcommon tree. So we all go back to a single, original group of people that lived in Africa about200,000 years ago. And so that was very exciting because there were other ideas. One idea was that

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in fact that people that lived in Africa, in Europe, and in Asia had each lived in those continents formany millions of years and had, each population had become human independently. If that weretrue, then we would have expected the mitochondrial DNA lineages to be like a set of three palmtrees, with very, very long stalks and then a bushy top, where, which would correspond to the peoplein Europe, Asia, or Africa. But rather than a cluster of palm trees, which would have given this so-called multiregional concept of human origins, we found that in fact it was more like a bush, withshort branches and then with the bushy part sticking out from the main core. So that implied that infact all the different populations were related to each other through a recent common ancestor, andthat common ancestor clearly was in Africa.

Cotton plantsJim Watsoncotton plants engineered to be pest resistant

Well you've got two cotton plants, one has the gene for the BT protein, and the bollworm can't growon it. Some were put on this plant when it was smaller and you've got a couple of spots where theycaused some necrosis, but if there was no gene this is what it would look like. In the old days, that iswhen pesticides became available and when I was Chairman of President Kennedy's committee onthe boll weevil, or cotton insects, they'd spray ten times a year, just to try and keep the plants lookinglike this. But I doubt they looked as good as this, this is a perfect plant. And you know, it's sort of wellPrince Charles would say nature is great, but it's only because you've intervened. If you left it alonethis is what the poor farmer has and after it no cotton. So you know, it's a case where you're tamingnature to man's needs. Now of course not the insects' needs, if you wanted an insect-dominatedworld this is it. But this is, you know, rational man's dominated world and to me this is the futureand a lot of farmers will have better lives, particularly the poor farmers.

Directing our evolutionJim Watsonour responsibility to direct our own evolution

Eugenics is sort of self-directing your evolution, and the message I have is that individuals shoulddirect the evolution of their descendants, don't let a State do it. I think it would be irresponsible notto direct your evolution if you could, in the sense that you could have a healthy child versus anunhealthy child, I think it's irresponsible not to try and direct the evolution to produce a humanbeing who will be an asset to the world as well as to himself.

American perspectiveJim Watsonan overview of eugenics in the USA

Here's a little pamphlet by Charles Davenport published in 1910 called “Eugenics” and it waspublished for the YMCA Health League, and the first chapter is called “Fit and Unfit Matings.” Andthis is what Davenport called the “Science of Human Improvement by Better Breeding.” The idea wasthat if someone had mental disease in the family then you didn't want to marry into it. So people hidmental disease. You wouldn't want to admit that your aunt was a bit wacky. The thing was the

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burden of the feebleminded, who’s going to take care of them and people began to try and look athow feeblemindedness moves through families and at Cold Spring Harbor they set up the EugenicsRecord station [sic, Office] just to do masses of pedigrees. They were going to prevent the unfit fromtaking over the world. Here's an apparent five generation family where feeble mindedness just runsthrough the thing and you can see here F, F, F, F, F, F, and I wonder how many of these kids went toschool, under what conditions and how they were tested and who said this. But this was the sort ofthing that affected social policy, the legislator who really didn't understand the science or thestrength of it passed laws to sterilize the unfit. Incredibly, as a result of Davenport's eugenics, 40,000women were forcibly sterilized in the United States.

Need for an RNA templateJim WatsonDNA makes RNA makes protein

Of course Francis and I talked about protein synthesis and there was evidence that protein synthesisoccurred in the cytoplasm of cells, not in the nucleus, so protein synthesis occurred removed fromthe chromosomes, or seemingly removed from the chromosomes. It occurred on particles whichcontained RNA. So I thought there must be some system by which the information is transferredfrom DNA to RNA and then RNA provides the information, is the direct template for proteinsynthesis. I wrote that on a little piece of paper and taped it above my desk.

Pre-WWII German eugenicsJim Watsoneugenics in Nazi Germany

This was the bigger version of the Eugenics Record Office at Cold Spring Harbor. The building wasbuilt with money from the Rockefeller Foundation in 1927, when eugenics was generally thought tobe a good thing. And the German geneticists thought it was a good thing and you know, had aproposed program of sterilization for a large number of genetic conditions. And, but it wasn't votedin, they couldn't get it through the German democracy at the time. But the moment Hitler came intopower a eugenics law was passed within a month, which prescribed sterilization for a large numberof conditions including for say, being schizophrenic and in a mental hospital. So very soonafterwards they started a program of sterilization which went on until the war started, with about600,000 people sterilized, it was a very thorough program and they had records on all these people.

None of us are perfectJim Watsonhuman imperfections and genetic enhancement

None of us are perfect. And if you could make us more perfect, why not. You know, and when we saywe're imperfect we mean we annoy people or we don't quite live up to what people expect of us, youknow, we can't kick a football right or you can't throw a ball or anything like this. So we spend somuch time trying to get better schools that I think any way you can enhance your future, fine. Andenhancing it through directing our script I think it's far off and it will be very difficult, and not assimple as people would think.

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Playing GodJim Watsonmiracles from knowledge, not prayer

People often ask me why do we want to “play God,” and my answer is simple, if we don't play God, nomiracles will occur and miracles occur through science. You know, there's virtually no polio becauseof science, and you know, there's no ulcers because of medicine and understanding, you know, nowthere's the bacteria down there, gives you ulcers, you know, we can cure it. So it's knowledge,miracles come from knowledge, not prayer.

DNA: the secret of lifeJim Watsonon his and Francis Crick's gigantic breakthrough

It didn't take long to sort of think, well, this is the answer. So you know, we probably came over hereby 12:30, I think we probably, you know, wanted to have a drink, you know, that, you know fromnowhere we suddenly had the answer. So we'd found the secret of life and Francis, you know,couldn't contain himself and, you know, you wanted to pinch yourself, it was so good. So you know,the people didn't know what we'd done, but it was hard to contain the fact that, you know, maybe wehad a gigantic breakthrough.

Reactions to imperfectionsJim Watsonshould we correct natural genetic imperfections?

The problem of the unfit is that there's constant variability generated by the imperfect replication ofDNA, that we're all the products of evolution and evolution wouldn't occur unless there's newvariability. So humans are born with mistakes in their, their book of life, and these mistakes are oftenvery debilitating. And Hitler thought, well we'll kill these people, and that's what effectively, youknow, he didn't want to see imperfect people. Now we like to get the knowledge to in some way, youknow, understand why they're imperfect and hopefully cure some, or get the knowledge by whichwomen might have the choice as to whether they will have an imperfect child.

The aimJim Watsonthe aim of the Human Genome Project

Discussions had been going on for about a year as to whether we should work out the completegenetic message in humans, it was called the Human Genome Project. And on the 24 differenthuman chromosomes there are some three billion bases, and the aim of the project was, get the orderof each of these bases along the chromosomes. And the people who first proposed it weretechnologists, they wanted to sequence bigger and bigger DNA molecules, they were talking abouthaving machines do it. And one thought, well they're going too fast, why don't you do a bacteria first.At that time we had some viral DNAs, we had about a hundred thousand, but we had to go, when you

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talked about humans it was ten thousand times bigger project.

The answerJim Watsonworking out the structure of DNAThe Cavendish shop was to build us some tin models and that took too long and, you know, finally indesperation I made some out of cardboard. I began moving them around and I wanted anarrangement, you know, where I had a big and a small molecule and so how did you do it? Somehowyou had to form link bonds, so here is A and here's T, and I wanted this hydrogen to point directly atthis nitrogen so I had something like this - Oh! So then I went to the pair. I wanted this nitrogen topoint to this one and it went like this - oh! They look the same, and you can put one right on top ofthe other. We knew if we could just, you know, even if we go up to the ceiling we were building a tinyfraction of a molecule. 100 million of these base pairs in one molecule, all fitting into this wonderfulsymmetry which we saw the morning of February 28th 1953.

Using family treesBarbara Weberidentifying and tracking genetic markers using family trees

So this is a family tree, drawn in a pretty standard way in the way that we do for these geneticstudies. And what’s on here is women in circles and men who are the squares –we always joke aboutthat! The women are colored in black if they have breast cancer, so you can see in this family thereare 1, 2, 3, 4, 5, 6, women who have breast cancer. And the lines tell you their relationship to eachother. So these are two sisters with breast cancer and their mother with breast cancer, her sister withbreast cancer, so these two are sisters and that’s the aunt of those two. And then her two childrenalso had breast cancer. The numbers underneath them are the genetic markers that we were using,and this was actually the pedigree that came into my mind as I was talking to Vicki that day in clinic.This is Vicki here, this is her sister who had died before I met them, and this is her sister Denise. Andwhat you can see, just sort of generally looking, is that what we had been doing was circling themarkers that went with the disease in this family and they're all the same – you probably can't seethem but this …all these people who have breast cancer have the same series of markers that areoutlined here. And what I could see clearly in my head was that Denise – that’s her – didn’t havethem.

Cancer genesMike Wiglerdescribing tumor suppressors and oncogenes

Cancer genes are roughly divided into two camps, oncogenes and tumor suppressors. The oncogenesare genes that sort of become super-genes, they drive the cancer cell. The tumor suppressor genesare the normal genes that we carry that are… whose function is to restrain the development andevolution of the cancers. They have other functions as well, but if they're lost then you're prone todevelop cancer. The oncogenes tend to be… we don't know all the oncogenes by a long shot. The onesthat we do know about, many of those are in fact enzymes.

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Clue: X-ray diffractionMaurice Wilkinsthe X-ray diffraction picture that revealed the helix

It was very exciting to see all the spots on the photographic film which were fairly sharp spots. Itshowed this sort of X-type - "oxo" type, of cross-pattern - which was an indication of a helix.

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