Dna microinjection
- 2. INTRODUCTION • DNA Microinjection is one of the method used for production of transgenic animals. • Organism containing integrated sequences of cloned DNA, transferred using techniques of genetic engineering are called transgenic animals. • This method allows an early integration of the transgene into the host DNA, which is important to ensure that transgenic DNA is apparent in all of the host.
- 3. PROCEDURE After 12 hrs of fertilization, fertilized oocytes are flushed from oviduct in a form of droplet in a Petridish. Super ovulated females are mated with male and then sacrificed Female animal is super ovulated by administering FSH and hCG hormones through i.v. Cloning of DNA into a vector as a plasmid Isolation of desired genes
- 4. Transgenic pups The injected embryos are subsequently transferred into the oviducts of pseudo pregnant female animal Transgenic DNA is physically microinjected into the pronucleus of a fertilized egg
- 6. ADVANTAGES • This technique is simple. • It is applicable to wide range of species. • Most widely used method.
- 7. DISADVANTAGES • Time consuming procedure. • Low success rate. • Unpredictable effects due to random transgenic integration-gene is over expressed or under expressed. • Costly
- 8. REFERENCES: 1. Liu, C., Xie, W., Gui, C. & Du, Y. Pronuclear microinjection and oviduct transfer procedures for transgenic mouse production. Methods Mol. Biol. 1027, 217–232 (2013). 2. Singhal, M. & Kansara, N. Transgenic Animals: Production and Application. Pharm. Sci. 1, 12–22 (2010).