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PRESENTATION ON TRANSGENIC ANIMALS AND ITS APPLICATION.pptx

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PRESENTATION ON TRANSGENIC ANIMALS AND ITS APPLICATION.pptx

  1. 1. MAHALAKSHMI.R II M.Sc-MICROBIOLOGY 1 PRESENTATION ON TRANSGENIC ANIMALS AND ITS APPLICATION
  2. 2. TRANSGENESIS • Transgenesis : Phenomenon of introduction of exogenous DNA into genome to create and maintain stable heritable characters. • Transgene : Foreign DNA which is introduced. • Transgenic animal: One whose genome is altered in heritable manner by foreign DNA sequence introduction into genome of its zygote or embryo(GEO/GMO) with new genetic character. • Transgenesis is facilitated by liposomes,enzymes,plasmid vectors, viral vectors, pronuclear injection, protoplast fusion and DNA injection. HISTORY • 1973: First GMO was created by S.Cohen and H.Boyer. 2
  3. 3. • 1974: First transgenic animals mice by Rudolf Jaenisch (SV40 viral DNA inserted in a mouse genome). • 1980: First transgenic mice via microinjection. • 1982: worlds first expressing transgenic animal (Super mouse- inserting a human growth hormone gene in mouse genome)- offspring will be larger than parents. • 1985: First transgenic rabbits,sheep,pigs and cattle. • 1996- 5-July: DOLLY was female domestic sheep and first mammal cloned from an adult somatic cell by nuclear transfer – Sir lan Wilmut , Keith Campbell and colleagues. PRODUCTION/METHODOLOGY • STEP 1: Construction of transgene –made of three parts: Promoter, Gene to be expressed and Termination sequence. • STEP 2: Introduction of foreign gene into animal. 3
  4. 4. • STEP 3: Screening of transgenic positives: Progeny are screened by PCR to examine site of incorporation of gene. • Some will not express as it integrated into transcriptionally inactive site. • STEP 4: Further animal breeding is done to obtain maxima expression-Heterozygous offsprings are mated to form homozygous strains. RETROVIRAL VECTOR METHOD • Transfer of small piece of DNA carried by retroviruses. • Results in chimera-org consists of tissues or parts of diverse genetic constitution. 4
  5. 5. MICROINJECTION METHOD •Super ovulation of young virgin female mice of 4-5 weeks by administration of follicle stimulating hormone followed by HCG(2 days). •Superovulated mouse produces 30-35 eggs (not normal 5-10 eggs). •Female mice are mated with males and fertilized eggs are removed from fallopian tubes. •By micromanipulation using a microinjection needle & holding pipette DNA is injected into male pronucleus. •Eggs with transgenes are kept overnight in incubator to develop two cell stage. •Eggs are implanted microsurgically to pesudopregnant mother which is mated previous night with vasectimized male. •Foster mother delivers pups after 3 weeks of implantation. 5
  6. 6. EMBRYONIC STEMCELL METHOD •Cells are taken from inner cell mass of blastocyst of developing embryo- proliferate in cell culture-pleuripotent ES cells. •It involves-introduction of foreign DNA into ES cells. And this in culture allows for gene manipulation using electroporation/microinjection. •Desired GE cells –identified by selection process using marker gene (thymidinekinase,enzymes- dihydrofolate reductase and neomycin phosphotransferase) and PCR. •Transfected cells cultured-introduced into blastocyst and then implanted into pesudopregnant female mouse. •Transgenic founder mice is produced. 6
  7. 7. EXAMPLES OF TRANSGENIC ANIMALS • TRANSGENIC MICE: Used as animals models for study of human diseases and for production of therapeutic agents. HUMAN MOUSE: Transgenic mice with human immune system produced-human mice. • Choose Mice with SCID and human thymus from aborted fetus–transplanted under capsule membrane of mouse kidney. • Human lymph node on opposite kidney of mouse. After week, immature immune cells from human fetus injected into mouse tail vein. 7
  8. 8. • These lymphocytes enters thymus under kidney and mature to T- lymphocytes. • This will enters circulation and lymph node. • Multiply to form full-pledged functional immune system. After 2 weeks ,Transplant mice display human immune system. APPLICATION: • Boon for immunologists for working on AIDS for possible development of AIDS vaccine . ALZHEIMER’S MOUSE: • Transgenic mice were developed by introducing amyloid precursor gene into fertilized egg cells of mice. 8
  9. 9. • Synthesis of human amyloid protein and plaques accumulation in mice brain. APPLICATION: • Understanding pathological basis of disease. ONCOMOUSE: • I-developed for breast cancer. • Introduction of chimeric DNA that contains c-myc gene & sections of MMT virus into fertilized egg cells of mouse. • Breast cancer in adult female mice and trait passed to offspring. APPLICATION: • Understanding of cancer and cancer therapy. 9
  10. 10. KNOCKOUT MOUSE FOR ALLERGY: • Developed by removing gene encoding for receptor protein so Ab cannot binds to cells due to lack of receptors and it is unaffected by allergic reactions. KNOCKOUT MOUSE WITH MEMORY LOSS AND RETINITIS PIGMENTOSA: • Develops mice with lack of hippocampus so that it lacks ability to remember. • Inactivation of mouse rhodopsin gene, rod cells in retina gets degenerated which is comparable to human retinitis pigmentosa. 10
  11. 11. TRANSGENIC CATTLE • Transgenic cow – over expressed casein transgene – milk with high content of casein. • Lactase transgene is introduced and expressed in mammary gland- milk free from lactose will be secreted. • Creation of transgenic cattle with improved Resistance to viral,bacterial and parasitic Diseases. TRANSGENIC SHEEP WITH INCREASED WOOL PRODUCTION: • Development of transgenic sheep has bacterial genes for synthesis of cysteine. 11
  12. 12. • Two enzymes synthesized by transgenes –traps H2S liberated in intestine to produce cysteine. TRANSGENIC PIGS: • It can produce human haemoglobin and separated by pig haemoglobin by AT. • Used for organ transplantation by harvesting human organs in it. • Because physiology of pig is close to humans, so organ transplant from transgenic pigs into human showed successful result. TRANSGENIC FISH: • SUPER FISH • Developed with increasing in their growth & size. • Introduction of growth hormone transgene by microinjection method. 12
  13. 13. • Fertilized eggs with inserted transgene are incubated in temperature regulated tanks. • Transgenic salmon fish is 10-12 times larger than normal fishes. GLO FISH: • Genetically modified zebra fish ,produce by integrating FP gene from jelly fish into fish embryo. DOLLY: First mammal clone developed by Wilmut & Campbell in 1997. • Sheep with mother and no father. TRANSGENIC MOSQUITOES: • Genes coding for endotoxin of P.vivax Introduced into mosquito zygote. 13
  14. 14. • Produces toxin of malarial parasites in their saliva and inject toxin to human blood through sucking blood. • Human immune system produce Ab against toxin and resist malarial parasites. TRANSGENIC MONKEY AND RABBIT: • ANDi –first transgenic monkey in 2000, inserted DNA spelled backward and insertion of GFP into its genome. • Proves transgenic primates can be created and express foreign gene delivered into their genome. • ALBA, inserted with enhanced GFP in 2000. IMPORTANCE OF TRANSGENIC ANIMALS: • Medical-disease model,pharma use, organ transplantation. • Agricultural- disease resistant animals, improving quality and quantity of milk,egg,wool production, breeding. • Industrial-toxicity sensitive transgenic animals to test chemicals. Produce proteins and enzymes for industrial reaction. 14
  15. 15. • ADVANTAGES: • Increased growth rate. • Improved disease resistance. • Increased muscle mass. • Improved nutritional quality. • Improved wool quality. • DISADVANTAGE: • Inserted gene has multiple functions. • Breeding issues. • Mutagenesis • Low survival rate. RECENT TRENDS: • Transgenic goats to produce human breast milk. • Bioluminescent transgenic mouse for study of mammary gland tumor development. 15
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  17. 17. THANK YOU 17

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