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8/2/2019 Transgenic Technology- Lecture #1 & 2 (MKM 05.01.2012)
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Transgenic Technology-
Introduction
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What is Transgenic Technology or Transgenesis?
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A transgenic organism
is a novel organism, whose genome contains therDNA and showed an altered trait or characteristic featurethat is economically or commercially desirable.
A technology/process used to make geneticallyengineered (GE) or genetically modified (GM) organism
A transgene
is a novel or recombinant DNA ( rDNA ) piece madefor the purpose of expression of a gene of interest (GOI) and trans ferred into the target host species .
Transgenic organisms are also created for basic research
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Horizontal gene transfer
Vertical gene transfer
What is new / special in transgenic technology ?
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Coding DNA sequence (CDS)Promoter Terminator
Essential DNA elements of a typical transgene for expression
Because the DNA of all organisms is chemically identical ; genes and
gene parts can be interchanged
All the three regions can be modified in the laboratory to control theabundance and activity of the gene product (RNA/protein/enzyme).
------ where , when , to what extent the gene will be expressed?
(Part of a vector used for transformation )
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TransgenicTechnology
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Applications of transgenic technology
In molecular biotechnology:
Developing commercially important product / organism
In functional genomics:
Identifying the functions of genes in the organism
During last 20 years or so, we have already witnessedthe power of this modern technology
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When the genome sequence is known, how do weidentify the function of gene?
Transgenic or reverse genetic approach
Genotype
Phenotype
Classical Genetics Reverse Genetics
Genotype
PhenotypeWild type & mutants
Alleles & genes
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Analysis of genetic mutations
(loss-of-function and gain-of-function )
The functions of the genes established
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Genome sequence data(Genotype )
Bioinformatics analysis to
predict the structures andfunctions of genes
Experimental approaches toestablish the functions of genes
(Phenotype )
Site-directed mutagenesis/ gene knockout / gene targeting(Not well established in plant system )
Insertional mutagenesisby T-DNA / transposon(Transgenic approach)
Gene silencing / RNAinterference
(Transgenic approach)
Wild type or mutant gene(over)-expression invarious host systems
(Transgenic approach)
Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions
Expression of Reporter Gene / Activation-tagging cassette
(Transgenic approach)
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Genome sequence data(Genotype )
Bioinformatics analysis to
predict the structures andfunctions of genes
Experimental approaches toestablish the functions of genes
(Phenotype )
Site-directed mutagenesis/ gene knockout / gene targeting(Not well established in plant system )
Insertional mutagenesisby T-DNA / transposon(Transgenic approach)
Gene silencing / RNAinterference
(Transgenic approach)
Wild type or mutant gene(over)-expression invarious host systems
(Transgenic approach)
Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions
Expression of Reporter Gene / Activation-tagging cassette
(Transgenic approach)
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Genome sequence data(Genotype )
Bioinformatics analysis to
predict the structures andfunctions of genes
Experimental approaches toestablish the functions of genes
(Phenotype )
Site-directed mutagenesis/ gene knockout / gene targeting(Not well established in plant system )
Insertional mutagenesisby T-DNA / transposon(Transgenic approach)
Gene silencing / RNAinterference
(Transgenic approach)
Wild type or mutant gene(over)-expression invarious host systems
(Transgenic approach)
Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions
Expression of Reporter Gene / Activation-tagging cassette
(Transgenic approach)
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Genome sequence data(Genotype )
Bioinformatics analysis to
predict the structures andfunctions of genes
Experimental approaches toestablish the functions of genes
(Phenotype )
Site-directed mutagenesis/ gene knockout / gene targeting(Not well established in plant system )
Insertional mutagenesisby T-DNA / transposon(Transgenic approach)
Gene silencing / RNAinterference
(Transgenic approach)
Wild type or mutant gene(over)-expression invarious host systems
(Transgenic approach)
Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions
Expression of Reporter Gene / Activation-tagging cassette
(Transgenic approach)
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Genome sequence data(Genotype )
Bioinformatics analysis topredict the structures and
functions of genes
Experimental approaches toestablish the functions of genes
(Phenotype )
Site-directed mutagenesis/ gene knockout / gene targeting(Not well established in plant system )
Insertional mutagenesisby T-DNA / transposon(Transgenic approach)
Gene silencing / RNAinterference
(Transgenic approach)
Wild type or mutant gene(over)-expression invarious host systems
(Transgenic approach)
Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions
Expression of Reporter Gene / Activation-tagging cassette
(Transgenic approach)
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Genome sequence data(Genotype )
Bioinformatics analysis topredict the structures and
functions of genes
Experimental approaches toestablish the functions of genes
(Phenotype )
Site-directed mutagenesis/ gene knockout / gene targeting(Not well established in plant system )
Insertional mutagenesisby T-DNA / transposon(Transgenic approach)
Gene silencing / RNAinterference
(Transgenic approach)
Wild type or mutant gene(over)-expression invarious host systems
(Transgenic approach)
Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions
Expression of Reporter Gene / Activation-tagging cassette
(Transgenic approach)
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Gain-of-function mutants:
(Over)-expression of the gene of interest (GOI) > New biochemicalfunction
Expression of marker/reporter gene cassette > Endogenous GeneTrap or Gene Tag
Expression of the activation-tagging cassette > Activation of theendogenous quiescent gene
Overview: Discovery of genes functions in post-genomic eraby transgenic routes to generate altered phenotypes
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Gain-of-function mutants:
(Over)-expression of the gene of interest (GOI) > New biochemicalfunction
Expression of marker/reporter gene cassette > Endogenous GeneTrap or Gene Tag
Expression of the activation-tagging cassette > Activation of theendogenous quiescent gene
Loss-of-function mutants:
Random integration of T-DNA or transposon-tagging cassette >insertional inactivation of unknown gene
Expression of specific endogenous gene-silencing cassette >siRNA-induced gene silencing or Gene knockdown
Targeted Gene Mutagenesis > Specific Gene Targeting or GeneKnockout (not well established in plant system )
Overview : Discovery of genes functions in post-genomic eraby transgenic routes to generate altered phenotypes
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What are the applications of transgenic research
in functional genomics ?
Identification of the function of genes
Isolation of genes or gene parts
Isolation of new promoter or other regulatory elements
Ultimate goal of functional genomics is to know / establishthe functions of all the genes and /or regulatory elements inan organism and to utilize them in molecular biotechnology
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Lecture # 1 delivered up to this slide on 04.01.2012
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Applications of transgenic research
In molecular biotechnology (using plant and animal):
Introgression of a gene of interest (GOI) to develop newor novel cultivar/livestock
Metabolic engineering to improve the existing elitevarieties/livestock
Enhancement of crop, meat, milk yield/ reduction of yield loss(quantity improvement )
Fruit, vegetable, meat, milk, egg and other food and feed
items for better post-harvest storage life, nutrition and health(quality improvement )
Large-scale production of industrial raw materials
Molecular farming for nutraceuticals and pharmaceuticals
S l d d l i h f M l l Bi h l /
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Selected developments in the areas of Molecular Biotechnology /Transgenic Technology
S l d d l i h f M l l Bi h l /
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Selected developments in the areas of Molecular Biotechnology /Transgenic Technology (Contd..)
Th F t t d f t
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The Farm- past, present and future
By Alexis Rockman
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George Bernard Shaw
You see things; and you say, 'Why?'But I dream things that never were;and I say, "Why not?"
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Transgenic Technology: Solutions to global problems?
Food shortage
Malnutrition
Diagnosis, prevention and treatment of disease & cancer
Healthcare
Industrial feedstock/raw materials
Renewable energy
Waste management / Environmental clean-up
Preservation of germplasms/ Conservation of biodiversity
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Transgenic Technology: Concerns and consequences !
Are GE/GM organisms harmful to other or to the environment?
Will the GE/GM organisms reduce the genetic diversity?
What are the ethical issues about GE/GM human being?
Will the financial support for Transgenic Research hamperother technological development?
Are the benefits of Transgenic Technology only for big
multinational companies/wealthy nations?
Are the patents/IPRs affect the free exchange of ideasamongst the scientists globally?
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Lecture # 2 delivered up to this slide on 05.1.2012