VBC-321 Animal Biotechnology Animal Cloning Cloning in many situations, highly desirable since this allows Indefinite multiplication of an elite desirable genotype without the risk of segregation and recombination during meiosis, which must
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Presentation Transcript
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VBC-321 Animal Biotechnology Animal Cloning<br>
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Cloning in many situations, highly desirable since this allows
Indefinite multiplication of an elite desirable genotype without the risk of segregation and recombination during meiosis, which must precede sexual reproduction
holds a great promise in genetic research, and impact of epigenetic changes
make it feasible to target transgenes in livestock by nuclear transfer from transgenic cell populations developed in vitro into enucleated oocytes to recover transgenic animals<br>
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TRANSGENIC ANIMALS Gordon and Ruddle used the term ‘Transgenic’ for the first time in 1982 to describe genetically modified animals harboring foreign genes within their genome
main objective in the generation of transgenic animals to guarantee that all the somatic cells and germ line cells carry the foreign gene
Production of transgenic animals involves a series of steps, starting with cloning of the gene of interest and preparation of DNA samples
both cloned and chemically synthesized DNA fragments can be used for microinjection
Microinjected embryos are usually transferred to the recipients after a short period of in vitro culture<br>
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Several methods are available for introduction of DNA into an animal genome. These include
DNA- microinjection into pronuclear stage of embryos
Use of retro viruses<br>
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STEPS IN PRODUCTION OF TRANSGENIC ANIMALS BY DNA MICRO INJECTION Females are superovulated and mated/ inseminated
Pronuclear stage – eggs are recovered after 12 hrs
Pronuclear transgene – DNA construct is injected into the male pronuclei by using micromanipulator
Embryos after microinjection are transferred to recipients and allowed to develop to term
Integration of the foreign gene in the new born animals could be confirmed by techniques like
Polymerase chain reaction
Southern hybridization
In situ Hybridization<br>
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APPLICATIONS OF TRANSGENIC ANIMALS To improve economic traits of farm animal like growth promotion, carcass improvement and milk production
To develop disease resistant animals
To produce a variety of biologicals and pharmaceuticals
To understand in vivo gene regulation<br>
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USES OF TRANSGENIC ANIMALS Basic research
Knockout mice for determining the function of a gene
To study effects of gene products,biochemical pathways, alternative (compensatory) pathways, and developmental pathways
To recreate human diseases in animals to establish models to test the beneficial effects of drugs or gene therapy
Knockout mice for genetic disease models<br>
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Production of useful proteins
Naked human Hb from pigs
Human lactoferrin in cows’ milk
Alpha-1-antitrypsin in sheep
HGH in mouse urine (uroplakinpromoters)
Human antibodies in mice (H and Lchain tgenics à hybridomas)
Tissue plasminogen activator (TPA)in goats
Human antithrombin III in goats
Malaria antigens in goats(vaccine)
Alpha-glucosidase in rabbits(Pompe’s disease<br>
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Bioreactor Genes transfered into animal with aview to obtain a large scale production of protein encoded by these genes in milk, urine, blood of such animal
These animals are bioreactor and approach is called molecular farming or gene farming<br>
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BIOPHARMING Biopharming is known as the production of pharmaceutical proteins using genetically engineered plants
promise to produce large and low-cost supplies of pharmaceutical drugs which includes vaccines for infectious diseases and therapeutic proteins for treatment of cancer and heart diseases
The proteins produced by the genetically engineered plants are called as Plant-made pharmaceuticals (PMPs)
The most common biopharm crops grown in US field trials are corn, tobacco and rice
Other crops being studied include alfalfa, potato, soybean, sugarcaen and tomato<br>
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NEED FOR BIOPHARMING Many pharmaceutical drugs have been produced in sterile fermenters by mammalian cells or using genetically engineered microorganisms. As the construction of huge fermentation plants incurs huge capital cost, the production costs also very high
Another method of production of biopharmaceuticals is to extract it from animal and human tissues such as insulin from pig and cow pancreas or blood proteins from human blood. But these procedures carry the risk of transmitting infectious diseases to humans
As there is advance in the techniques of manipulating the plant genome over the past several years, plants can be used to produce a wide range of important proteins
It is hoped that this will result in therapeutic products at a price significantly cheaper than through current methods of production<br>
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CRITERIA TO SELECT A PLANT FOR BIOPHARM PRODUCTION It should be easily engineered
It should be capable of production of high levels of proteins
Appropriate technique should be available to extract the proteins from plant tissues
Ideally the host plant should be non-food crop or the food crop should be completely sterile to avoid cross-pollination with near by field crops<br>
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RISKS OF BIOPHARMS Pollen from plants engineered to produce pharmaceuticals may fertilize nearby food or feed crops of the same species. If this occurs, the pharmaceutical may be produced in seed of the neighboring crop, with potentially negative effects on human or animal consumers of the seed
The introduced gene or its product may have negative effects on the natural environment
Farm workers may be exposed to unhealthy levels of a biopharmaceutical by absorbing products from leaves through their skin, inhaling pollen, or breathing in dust at harvest
Unexpected toxins or residues of pesticides used on the crop may contaminate the final drug product<br>
