Transgenic Technology- Lecture #1 & 2 (MKM 05.01.2012)

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


Reverse genetics approach:Transgenic or reverse genetic approach for discovery of genes functions

Expression of Reporter Gene / Activation-tagging cassette



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(Phenotype )











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(Phenotype )











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(Phenotype )











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Bioinformatics analysis topredict the structures and

functions of genes


(Phenotype )











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Bioinformatics analysis topredict the structures and

functions of genes


(Phenotype )











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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

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Transgenic Technology- Lecture #1 & 2 (MKM 05.01.2012)

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