Transgenic Animals: Objectives of Gene Transfer with Different Transfection Methods! Nature. Thus, one method to create a transgenic animal involves gene transfer (transgene incorporated into the genome). The Embryonic Stem Cell Method (Method "1") Embryonic stem cells (ES cells) are harvested from the inner cell mass of mouse blastocysts. Gene delivery is the process of introducing foreign gene delivery, such as DNA or RNA, into host cells. It was not until recent progress of the retrovirus vector system that high efficiency of this gene delivery system was demonstrated in the production of transgenic pigs, cattle, and primates (13–16). Cell. Retroviral vectors carrying foreign nucleic acid and having a Feline Leukaemia Virus B env gene product can be introduced into the embryos of sheep and pigs. Gene transfer with retroviral-vector assistance. This paper. For instance the very efficient DNA transfer was attained in Swine which was up to 80% of the offspring were transgenic (Lavitrano et al., 2002) while in the case of bovine this method had limited achievement to produce transgenic cattle (Schellander et al 1995) therefore, sperm-mediated gene transfer has not been widely adopted for making transgenic animals unlike the other technique. In transgenic animals, however, the foreign gene is transferred indiscriminately into all cells, including germ line cells. Two methods of producing transgenic mice are widely used: transforming embryonic stem cells (ES cells) growing in tissue culture with the desired DNA; injecting the desired gene into the pronucleus of a fertilized mouse egg. The retroviral vector was injected into laid quail embryos at the blastodermal stage, and the embryos were incubated to hatch to produce G(0) transgenic quails. Replication-defective retroviral vectors are currently the most efficient gene transfer method for the production of transgenic animals. We microinjected retrovirus into the seminiferous tubules of 5- to 10-day-old B6, C3H, BALB/C, and A mice (founder mice). PDF. Retrovirus vectors have been used for a variety of applications requiring gene transfer, including the production of transgenic animals. 1989 Oct 20;59(2):239-41. Transgenic animals 2. Transgenic animals mice for students 1. Transgenic animals are raised and mated. These keywords were added by machine and not by the authors. In all such cases, the transgene is delivered through an infectious particle and as part of a retrovirus genome. Transgenesis in Mice Retroviral Vector • Efficient mechanism of transgene integration • Transfer of genes < 8 kb possible • random insertion of the transgene • Retroviral contamination of the transgenic animal Transgenesis in Mice Retroviral Vector. In almost all cases, the retrovirus genome is replication defective because its genes have been replaced with the transgene. PDF. transplantation method, retroviral vector meth od, germ line stem cell mediated method to improve efficiency, gene targeting to improve accuracy, RNA interference-mediated gene silencing technology, zinc-finger nucleases gene targeting technique and induced pluripotent stem cell technology. A method for producing a transgenic animal comprising the steps of: transferring transgenic embryos, which were produced by transferring exogenous DNA to oocytes by injection of a retroviral vector, fertilizing the oocytes by intracytoplasmic sperm injection, and culturing said oocytes to the embryonic stage, to the oviducts of surrogate females; and producing a transgenic animal by parturition. Given the drawbacks of retroviral transgenesis, DNA microinjection in the pronucleus became the most widely employed method to create transgenic animals. 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