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KMID : 0900920010250020171
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Korean journal of Animal Reproduction
2001 Volume.25 No. 2 p.171 ~ p.180
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Gene Transfer into Chicken Embryos using Defective Retroviral Vectors Packaged with Vesicular Stomatitis Virus G Glycoprotein Envelopes
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Heo Young-Tae
Lee Hoon-Taek
Lee Young-Man
Kim Teo-An
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Abstract
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Compared to other gene transfer system, the advantages of retrovirus-mediated gene transfer are technical ease, efficient expression and genetic stability. Despite the high potency of the retrovirus vector system in gene transfer, one of the drawbacks is a difficulty in concentration of virus stock. To overcome this problem, we tested a new retrovirus vector system producing the progeny retrovirus particles encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein). The infectivity of this virus was not sacrificed by ultracentrifugal concentration and the host cell range extended from all mammalian to fish embryos. Virus titer after 1,000 x concentration was more than 10^{8} CFU/ mell on most of the target cell lines. We applied this pantropic viruses in transgenic chicken production by injecting the concentrated (100times) stock into subgerminal cavity of stage X chicken embryos. The survival rate of chicken embryos after injection was about 20% and gene integration rate in surviving embryos was scored almost 100%. Analyses of RT-PCR and fluorescence microscopy, however, showed no evidence of the transgene expression.
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KEYWORD
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Retrovirus vector, VSV G, Concentration of virus stock, Embryo injection
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