CRISPR-Cas9--Genome Editing (.pptx)

Yesterday, Today, and Tomorrow

March 31, 2016 11/24/2016Paradise Valley Community College1

As a small boy in the 1950s, I was more interested in catching bumblebees and riding my Schwinn than what was happening in the bigger world around me.

But that world eventually caught up with me, pulled me in, and opened my eyes to the amazing world of biology.

More than sixty years later, many spent in the study of chemistry and biology, this short video caught my attention and caused me to look back on those years.

(Start the video)

Today, I would like to take you on a journey back in time to give you a perspective as to why this Breakthrough of the Year is so important as we move into the future.

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Todays TopicsGenome Editing

Yesterday,Today,and TomorrowViruses

Immunology

Biotechnology

PatentsEthicsNobel PrizesBacteria11/24/2016Paradise Valley Community College2Critical Thinking

Nobel PrizesBacteriaVirusesImmunologyBiotechnologyPatentsEthicsCritical Thinking

On this journey, we will address these subjects.

It is impossible to give them the time they deserve but it is also imperative that we create the framework that leads us to Tomorrow.

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Photo 51

1928-

1916-2004

1905-2002Yesterday (1953-1962)Erwin ChargaffDiscovered the relationships between DNA bases, A, T, G, CRosalind Franklin & Maurice WilkinsDiscovered the basic structure of DNA by x-ray crystallographyJames Watson and Francis CrickBuilt the first accurate modelof a DNA molecule

1920-1958

1916-2004

1949

1962

1962

196211/24/2016Paradise Valley Community College3The Central Dogma of Molecular BiologyDNA makes RNA makes Protein.

3Our journey starts with this image. Photo 51, as designated by Rosalind Franklin, is an X-ray crystallographic image of purified DNA. The symmetry is apparent and would become the touchstone of all that would come.

Earlier in the 20th century, Edwin Chargaff established the relationship between DNA and the nucleic acids, A, T, G, and C. Well leave the chemistry for others to explain but note that he was awarded the Nobel Prize in 1949.

During the early 50s, Franklin and Wilkins discovered, via this x-ray image, the very basic structure of DNA.

James Watson and Francis Crick took this image and built the first model of DNA. This work also achieved Nobel Prize status in 1962. Note that that prize was awarded to Watson, Crick, and Wilkins. The rules of the award allow only three winners per prize. Franklin had died in 1957 of cancer.

This watershed moment eventually led to what is now called the Central Dogma of Molecular Biology, DNA makes RNA, makes Protein. Simply put, but soon to become much more complicated.

11/24/2016Paradise Valley Community CollegeChapter 6 - DNA Structure and Function

Yesterday (1970)11/24/2016Paradise Valley Community College4

Christian B. Anfinsen, Jr.1916-1995

The Central Dogma of Protein FoldingDNA dictates the amino acid sequenceand that determines protein functionality. 1970:Protein Folding of Ribonuclease

1972Christian AnfinsenStanford MooreWilliam Stein

In 1972, I was a graduate student at Indiana University working on protein chain initiation. Next to our obsessively intense and sedate lab was the lab of Carlos Miller, renowned for his work on plant hormones. He had a graduate student that was a good friend of mine, Carol Crafts.

Picture in your mind our ordered lab, a Sorval centrifuge running at speed, a scintillation counter clicking away in the background, and me grinding bean leaves to dust in liquid nitrogen.

The door flies open and Carol shouts out, Guess what! Daddy just won the Nobel Prize!

Daddy was Chris Anfinsen and he, along with Stanford Moore and William Stein had just won the 1972 prize, for their work on the structure of ribonuclease.

They were able to elucidate the folded structure of this enzyme, accurately determining the hydrogen-bonding and disulfide bridges that create the shape you see here. A more recent graphics is shown next to it.

Anfinsen was attributed with a second Dogma: DNA dictates the amino acid sequence and that determines proteinfunctionality. Put another way, the primary structure of a protein leads to its tertiary structure, i.e., functionality. DNA dictates the amino acid sequence and that determines protein functionality.

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ATTGGTAACCTTAAAATTCAGCAGTCGAYesterday (1978)1970:Discovery of EndonucleasesEnzymes capable of cleaving DNA at very specific locations11/24/2016Paradise Valley Community College5

1978Werner ArberHamilton SmithDaniel Nathans About 3,000 known todayfor 230 DNA sites(Source: www.promega.com. 2015.)Source: en.wikipedia.org/wiki/Restriction_enzyme. 2015.

Innate Immunity in Bacteria

Later in this decade, an enzyme, termed an endonuclease, was found in bacteria that cut DNA with high specificity.

The graphic on the left illustrates the tertiary structure of this enzyme with its active domains cutting the DNA in orange.

This enzyme cleaved DNA over six bases, breaking the DNA and leaving 4 unpaired bases exposed.

This work won the Nobel Prize for Arber, Smith, and Nathans, postulating an innate immune response in bacteria.

Today, we have identified about 3,000 such endonucleases cleaving about 230 sites on the DNA.

This discovery intimated that scientists could now insert bits of DNA into an organisms complement of DNA, the genome.

The excitement around this advance also created concern. The phrase Frankensteins Monster came up frequently in the press. Ethical questions led to this conference in which 140 scientists convened to create some rules.

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Yesterday (1975)1975:Monterey, CA: Asilomar Conference on Recombinant DNA140 ScientistsOutcomesGuidelines about how to isolate dangerous experimentsDetermination thatcloning and messingaround with dangerouspathogens should beoff-limitsModifying the humangerm line was nota worry.

Source: news.usc.edu, 2015.11/24/2016Paradise Valley Community College6

This group debated the concerns and came up with two major outcomes.

Guidelines were created to insure isolation of the experimental work and that defined pathogens would be considered off-limits for research.

It is important to point out that this research was too early to even consider using it when working with the human genome.

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Yesterday (1980)11/24/2016Paradise Valley Community College7Sticky endsSticky ends

DNA LigaseRecombinant DNA (rDNA)Native DNASticky ends

InsectResistant Gene

Restriction Endonuclease (EcoRI)

1980Paul BergWalter GilbertFrederick Sanger

Lets take a closer look at what the research was about and why it prompted the Asilomar Conference of 1979.

Here we see a short segment of a plant DNA and the repeating six bases are labeled. The endonuclease cleaves the DNA leaving the ends of the DNA sticky, missing their previously intact hydrogen bonds.

A gene designed to create insect resistance is added to the cleaved DNA of the plant. It, too, has been created with sticky ends complementary to the sticky ends of the cleaved DNA.

These sticky ends seek each other out and another enzyme, DNA ligase, binds the ends and re-establishes the DNA backbone.

This gene addition creates what is called recombinant DNA. Above it, for comparisons sake, is the original native plant DNA.

This research won the Nobel Prize for Berg, Gilbert, and Sanger in 1980.

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Yesterday (1987-2007)1987:Discovery of Repeating Sequences in BacteriaSo far, no sequence homologous to these has been found elsewhere in prokaryotes, and the biological significance of these sequences is not known.1993:Found in 20 bacteria and named SRSR2002:Name of CRISPR first applied.Clustered Regularly Interspaced Short Palindromic Repeats2005:CRISPR provides acquired resistance against viruses in prokaryotes.11/24/2016Paradise Valley Community College8Ishino, Y, et al. J Bacteriol. 1987 Dec; 169 (12): 542933.

Source: www.ncbi.nlm.nih.gov/. 2015.Barrangou, R., et al. (2007) Science 315:1709Jansen, R., et al. (2002). Mol. Microbiol. 43: 1565.Mojica, F. J. M., et al. (1993). Mol. Microbiol. 36: 244246.

A few years later in 1987, Ishino and his colleagues, published a paper showing a gene from E. Coli. I apologize for its readability, but I believe you can see the pattern repeating itself in these 300 nucleic acid bases.

Their conclusion? So far, no sequence homologous to these has been found elsewhere in prokaryotes, and the biological significance of these sequences is not known.

Further research led to a description of Ishinos repeating sequences and a name: Clustered Regularly Interspaced Short Palindromic Repeats or CRISPR for short.

Little more was known about it. In 2005, CRISPR was shown to provide acquired immunity against viruses in prokaryotes.

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2001Source: www.slideshare.net. 2015

2002Source: www.slideshare.net. 2015Yesterday and Today1990-2002:The Human Genome Project (HGP)The draft human genomesequence was publishedon February 15, 2001.The mouse genome wassequenced 13 months later.11/24/2016Paradise Valley Community College9Today, roughly 18,719 species have been sequenced. (Source: www.the-scientist.com. 2015)

Obviously, scientists had to learn more about the genome to continue further down this path.

Bacterial genomes were being sequenced at that time but now comes a big push to understand the human genome.

This Human Genome Project involved academic and private sector scientists. More than a decade passed.

In 2001, the draft human genome was elucidated and published in Nature. Thirteen months later, the mouse genome was known.

Today, in 2016, we have sequenced almost 20,000 species.

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Yesterday (1971)11/24/2016Paradise Valley Community College10

1993Kary MullisMichael Smith

The improvements made by Mullis allowed PCR to become a central technique in biochemistry and molecular biology.(Source: en.wikipedia.org/wiki/Kary_Mullis )First described by Kleppe & Khorana 1971:Polymerase Chain Reaction (PCR)

1971. Journal of Molecular Biology 56 (2): 341361.

Lets rewind the story a bit. The discovery and refinement of the polymerase chain reaction during the 70s and 80s allowed rapid genome sequencing.

The technique was first described by Kleppe and Khorana and later refined by Mullis and Smith for which the latter pair received the Nobel Prize in 1993.

Kary Mullis career was varied. From Ph.D. to fiction writer to bakery manager, Mullis then went back into science, working for a number of biotech companies in California.

He may have been the first to suggest the use of a heat-resistant polymerase in the PCR reaction. Regardless, a technique was now available that could amplify small amounts of DNA, essentially purifying it from unwanted cellular material.

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Genomics & ForensicsPCR quickly replicates copies of a particular DNA fragment for study. Primer nucleotides initiate the process.Polymerase Chain Reaction (PCR)Primers and enzymes rapidlygenerate many copies ofa DNA fragmentPrimers (short DNA strands)Free nucleotidesTaq DNA polymeraseTemperature cycling

11/24/2016Paradise Valley Community College11

11The PCR technique became the standard in both genomics and forensics.

The DNA fragment of interest is mixed with free nucleic acids, ligases, Taq polymerase, and primer DNA with incubation at both low and high temperatures.

The DNA fragment of interest might be metaphorically thought of as the needle in the haystack. PCR allowed scientists to amplify the DNA until it became the haystack.

11/24/2016Paradise Valley Community CollegeChapter 10 - Biotechnology

249,250,621

Yesterday and Today11/24/2016Paradise Valley Community College12

1249,250,6212243,199,3733198,022,4304191,154,2765180,915,2606171,115,0677159,138,6638146,364,0229141,213,43110135,534,74711135,006,51612133,851,89513115,169,87814107,349,54015102,531,3921690,354,7531781,195,2101878,077,2481959,128,9832063,025,5202148,129,8952251,304,566X155,270,560Y59,373,566690,472,424881,626,7001,062,541,9601,392,795,6901,539,159,7121,680,373,1431,815,907,8901,950,914,4062,084,766,3012,199,936,1792,307,285,7192,409,817,1112,500,171,8642,581,367,0742,659,444,3222,718,573,3052,781,598,8252,829,728,7202,881,033,2863,036,303,8463,095,693,981492,449,994Chr#Base Pairs Chr#Base Pairs mt16,569Cumulative Base Pairs Total base pairs in the human genome (draft)

Now back to the human genome and what we currently know, a code of many As, Ts, Gs, and Cs, and their proper places on the 46 chromosomes and mitochondria.

More than three billion letters define the genome. But lets look at what 3 billion looks like.

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Size of the Human Genome11/24/2016Paradise Valley Community College13AGTCCATTACGAAAATCGACTATCGAAGGGTAAAGGCTTATAAGCCATAGACATAGATAACTACCTTAGGAATATCAGTACGATTAAATGCCCATGAATCGAATTGGACCATAGCTAAGATCAGATCTAGTATCGAATGCTTATAGCCCATGGATACGATCAGATCAGATACGATAGTACATGCAATGGATCACCTAGATGGATCGATTAGGAATCCACCCATGTGGCATACCTAATTTGAAGAAAGACTACCTTAGGAATATCAGTACGATTAAATGGCCCATGAATCGAATTGGACGAATCGAATGCTTATAGCCCATGGATAAt the rate these bases are appearing, it would take 5 years of watching this to see the complete human genome.

Self-explanatory13

Yesterday & Today Drives TomorrowFeeding the WorldEradicating DiseaseImproving Quality of LifeIncreasing the Life-SpanPreventing Disease Outbreaks

Yesterday & TodayCentral Dogmas:DNA makes RNA makes PROTEINProteins tertiary structure conveys functionalityEndonucleases:Accurate & specific DNA cleavageRecombinant DNA alters native DNAAdvances driven by technology:Polymerase Chain Reaction (PCR) DNA sequencing creates the roadmap11/24/2016Paradise Valley Community College14

Lets take a moment to summarize.

Early landmark work on the roles of DNA, RNA, and protein was established.

Protein functionality was found to be a result of its tertiary structure.

Enzymes were discovered that specifically cleaved DNA allowing for the creation of recombinant DNA.

Technology gave us a means of rapidly sequencing the genome, hence, creating a roadmap for alterations.

All for the purpose of what? Answering questions that have plagued humankind well into the past. I cannot possibly cover all of these topics, so I will mention only a few as they bear upon Tomorrow.

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Source: seedtoday.com. 2016Feeding the World: YesterdayConventional BreedingCurrent cultivar crossed with wild type.Backcross offspring with cultivar.Meiotic crossovers slowly eliminate unwanted genes.Process is slow and costly.

X

XF2F1FnBeneficial gene

Unwanted genesBackcrossManyBackcrossesNewCultivarWild type

11/24/2016Paradise Valley Community College15CurrentCultivar

Source: modified from ucfant3145f09-06.wikispaces.com/Wilson's+Page. 2016.

15When I was a child in Iowa, we often drove through the farmlands to visit relatives. Corn, twice as tall as I was, came right up to the roadside. This field-post marker of corn with wings flashed by as we drove. This was my first exposure to the science of conventional breeding.

Current cultivars were genetically exposed to other cultivars that might improve the current crop.

Cross pollination of one with the other allowed incorporation of the beneficial gene along with other unwanted genes. A backcross to the current cultivar slowly eliminated the unwanted genes.

Many such backcrosses over many growing seasons eventually led to the desired hybrid. A slow and laborious process.

But, over many centuries, this is the result.


Feeding the World: TodayGene Modification (Cisgenesis)A gene may be modified and reinserted into an individualof the same species.Gene introduced bybacterium or DNA Gun.Plasmid inserts gene.Process requires one generation.

PlasmidNewCultivarCurrentCultivar

Plasmid

WildRelative11/24/2016Paradise Valley Community College16

16The discoveries and techniques of yesterday allow for gene modification today, typically referred to as cisgenesis and transgenesis.

Cisgenesis moves genes from one organism to another organism of the identical species.

Genes of the related wild type can be isolated and moved to the current cultivar using a bacterial vector (agrobacterium, for example) or by encapsulating the gene in gold nanoparticles and shooting these into the current cultivar.

In a single generation the beneficial gene is incorporated into the current cultivar.

Is this a GMO, a genetically modified organism, as defined by the FDA? Hold that thought.11/24/2016Paradise Valley Community CollegeChapter 10 - Biotechnology

Feeding the World: TodayGene Modification (Transgenesis)Genes inserted into a currentcultivar from different speciesDNA from Chinook SalmonAntifreeze gene from Ocean PoutOva from Atlantic SalmonRapid and cost-effective process

UnrelatedorganismsPlasmidNewCultivarCurrentCultivar

Plasmid


Source: modified from trylivingorganic.com. 2016

17Transgenesis differs from cisgenesis.

Genes are taken from an unrelated species, inserted into a bacterium or encapsulated in nanoparticles, to be vectored into the current cultivar, creating the new cultivar, again in a single generation.

As an example, lets look at how this is accomplished in the salmon some of us consume. Three distantly-related organisms are used. DNA segments of the Chinook Salmon and the antifreeze gene of the Ocean Pout are incorporated into the ova of Atlantic Salmon.

The result? This is classified as a GMO.


Yesterday, Today, and Tomorrow


This false-color electron micrograph shows viruses (bacteriophages) infecting a bacterium.

The viruses attach to the bacterium and inject their DNA into it.

What prevents a complete extinction of the bacterial species? Adaptive immunity.

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CRISPRAdaptive Immunity in Bacteria11/24/2016Paradise Valley Community College19Clustered Regularly Interspaced Short Palindromic Repeats

CRISPR

SSSSSSSCas9

CRISPR-Cas9 Complex Bacterial DNA

PlasmidBacterium

Bacteria respond to bacteriophage infection in this way.

The bacterium senses the presence of the foreign DNA and digests it as best it can. A small segment of the viral DNA is then incorporated into the genome of the bacterium near a gene locus called Cas9. This occurs with each new viral infection; spacers being inserted between each new piece of viral DNA.

The memory of past infections is now set and we have the genetic sequence referred to as CRISPR, clustered regularly interspersed palindromic repeats.

If viral infection recurs, then the bacterium again digests the viral DNA and finds a match within its CRISPR segment. This activates the Cas9 gene to produce a protein and the CRISPR segment to reproduce its spacer and viral DNA segment transcribed into RNA. The CRISPR-Cas9 complex is created. Lets take a closer look at this complex and its function.

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Adaptive Immunity in BacteriaGuide RNA (gRNA)Viral DNAMatchingSequences

Cas-9 ProteinSource: modified from origene.com. 2015.

Viral genome destroyed

Viral DNA


The Cas9 protein seeks out a region of viral DNA that matches the CRISPR RNA segment, the Guide RNA) imbedded in the Cas9 protein.

Molecular scissors snip the viral RNA and the viral genome is destroyed.

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UK, February, 2016:approval given for editing the human embryoChina, April, 2015:corrected beta thalassemia gene implanted in human embryo

DonorDNAThe Tools of Genetic EngineeringGuide RNA (gRNA)Genomic DNAMatchingSequences

RepairCas-9 ProteinSource: modified from origene.com. 2015.

Genomic DNA11/24/2016Paradise Valley Community College21

Targeted genome editing

Jennifer Doudnas lab at Berkeley discovered that donor DNA could be attached to the Guide RNA.

The CRISPR-Guide RNA could be engineered to create any sequence desired and then matched to any genomic DNA.

The genomic DNA is snipped, the donor DNA inserted, and the genomic DNA repaired. This is the model for CRISPR-Cas9 targeted genome editing.

In the last three years, this has been performed on bacteria, zebrafish, mice, and pigs. In April of 2015, Chinese scientists demonstrated that they could correct the HBB gene which causes beta thalassemia in a human embryo. A point of clarification should be made: these zygotes did not proceed through embryogenesis.

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Today11/24/2016Paradise Valley Community College22

Targeted Genome Editing

This short video summarizes the previous explanation.22

Today and TomorrowGene ModificationYeastNematodesDrosophila melanogasterAxolotls (knockouts)Xenopus tropicalisSilkworm (spider silk)Zebrafish (bi-alleic editing)Mice (DMD)Rats (germline)Pigs (biomedical models)Monkeys (disease models)Somatic Genome EditingMouse liverMouse lung

Correction of Genetic DisordersMouse cataractsMouse muscular dystrophy (DMD)Human cystic fibrosis (CFTR)Cultured intestinal stem cellsGenomic Screening ResearchCRISPRi (gene repression)CRISPRa (gene activation)Treating Infectious DiseasesRetroviruses (HIV)DiphtheriaAnthraxCholeraViral Hepatitis BSource: Yang, Mil Med Res. 2015; 2: 11. 2015.11/24/2016Paradise Valley Community College23

Here is a current list illustrating present and future use of the CRISPR-Cas9 complex.

Gene modification, somatic genome editing, correction of genetic disorders, genomic screening research, and treating of infectious diseases as seen in the following video.

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Tomorrow11/24/2016Paradise Valley Community College24Elimination of Malaria

Self-explanatory24

Pros & ConsBenefits (Today)Mutations reversed that cause blindnessCancer cells stopped from multiplyingCells made impervious to the HIV & Ebola virusesBread wheat rejects powdery mildewYeast altered to consume debris & generate ethanolCosts are low ($699 for complete kit)

Concerns (Tomorrow)If new genes wipe out malaria, what happens to bats (keystone species)?Off-Target EditingEugenics & designer babiesInvasive mutationsBio-weaponsCosts are low ($699 for complete kit)

11/24/2016Paradise Valley Community College25Source: www.wired.com/2015/07/crispr-dna-editin.g-2/. 2025.

The possible benefits are myriad. These are just a few examples. The last one is most important.

Research studies that previously cost thousands of dollars can now be preform in your kitchen.

The concerns that arise are just as pervasive.

he most important is, perhaps, that costs permit you to do this research in your kitchen.

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TomorrowDoudna, J. and Charpentier, E. June, 2012.CRISPR was used to edit purified DNA.(Patent dispute, pending final approval)

Zhang, F, and Church G. January 2013.CRISPR was used to edit human cells.(Patent dispute, pending final approval)11/24/2016Paradise Valley Community College26

Source: www.brandeis.edu. 2015

?

DoudnaCharpentier

ZhangChurch

The ground-breaking work was performed in two labs. Doudna and Charpentier at Berkeley won the Breakthrough Prize of $3 million for their work demonstrating that CRISPR could be used to edit purified DNA. Zhang and Church at the MIT/Broad Institute refined the earlier work showing that CRISPR could be used to edit human cells. Zhang applied for a patent first which was countered by Doudna. This dispute hangs fire right now. So, is this Nobel Prize research and who should be so awarded? Remember the rule: Only three awardees per prize.

In January 2013, Zhang's team published a paper in Science showing how Crispr-Cas9 edits genes in human and mouse cells. In the same issue, Harvard geneticist George Church edited human cells with Crispr too. Doudna's team reported success in human cells that month as well, though Zhang is quick to assert that his approach cuts and repairs DNA better.

That detail matters because Zhang had asked the Broad Institute and MIT, where he holds a joint appointment, to file for a patent on his behalf. Doudna had filed her patent applicationwhich was public informationseven months earlier. But the attorney filing for Zhang checked a box on the application marked accelerate and paid a fee, usually somewhere between $2,000 and $4,000. A series of emails followed between agents at the US Patent and Trademark Office and the Broad's patent attorneys, who argued that their claim was distinct. WIRED26

Washington, DC 3 December 2015Basic and Preclinical ResearchNeeded and should proceedClinical Use: SomaticProceed if only the individual is affectedClinical Use: GermlineIrresponsible to proceedNeed for an Ongoing ForumU.S. National Academy of ScienceU.S. National Academy of MedicineThe Royal Society (UK)The Chinese Academy of Sciences11/24/2016Paradise Valley Community College27Source: www8.nationalacademies.org/onpinews/newsitem.aspx?RecordID=12032015a. 2015

embryos

Recall the Asilomar Conference of 1975. Another such conference was just held in Washington, sponsored by the U.S. Academy of Science, the U.S. Academy of Medicine, the British Royal Society, and the Chinese Academy of Sciences.

Scientists created four binding resolutions. Basic and preclinical research should proceed, somatic clinical studies should proceed in affected individuals, germline studies would currently be irresponsible without government oversight and regulation, and ongoing committees are needed as data is developed and refined.

The third point is critical.

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In 1953, Jonas Salk was askedWho owns the patent on the polio vaccine?His response?Well, the people.There is no patent.Could you patentthe Sun?11/24/2016Paradise Valley Community College28

Source: timemarcheson.wordpress.com. 2016.

Source: tripod.com. 2016.

As a young boy, my mother took me to a clinic and I was given a cherry-flavored syrup to drink. That was the polio vaccine. Jonas Salk was responsible for the nearly complete elimination of the disease in the U.S.

He was asked, Who owns the patent on the polio vaccine?

His response?

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NOTThe EndThe BeginningStephan A GeorgeAdjunct Faculty, Life SciencesParadise Valley Community CollegeMarch, 2016

DNA

Able was I ere I saw elbado geese see god11/21/2015Paradise Valley Community College30

PalindromePalindrome

RNA

Pompeii, 79 A.D.The SATOR SquareRNA

Today and Tomorrow11/24/2016Paradise Valley Community College31One copy of the altered gene is inherited.Leads to a 50 percent chance of passing it on per generation.Elimination of Malaria(Non Gene Drive)Gene drives ensure both copies of the altered gene are inherited.Leads to a 100 percent chance of passing it on.Elimination of Malaria(Gene Drive)TodaYTomorrow

Washington, DC - December 3, 2015Sarah GrayBore a son with anencephaly, who suffered seizures for 6 days until he diedIf you have the skills and the knowledge to eliminate these diseases, then freakin' do it.11/24/2016Paradise Valley Community College32

Sarah Gray, Director of Marketing and Public Affairs, American Association of Tissue Banks Source: news.sciencemag.org/scientific-community/2015/12/inside-summit-human-gene-editing-reporter-s-notebook. 2015.

The science of regulation is more precarious and uncertain than the science of gene editing.11/24/2016Paradise Valley Community College33Barbara EvansProfessor of Law and George Butler Research ProfessorDirector, Center on Biotechnology & LawUniversity of Houston Law Center, Houston, Texas

Source: news.sciencemag.org/scientific-community/2015/12/inside-summit-human-gene-editing-reporter-s-notebook. 2015.

IPOs and suppliers11/24/2016Paradise Valley Community College34

addgene.orgaudentestx.com

editasmedicine.com

intelliatx.comcorvuspharma.com

reatapharma.com

syndax.com

origene.com