A highly efficient method for the site-specific integration of transfected plasmids into the genome of mammalian cells using purified retroviral integrase.
Using purified bovine leukemia virus (BLV) integrase with liposome, we developed a highly efficient method for the site-specific integration of plasmid vectors into the genome of cultured mammalian cells. The presence of the BLV integrase recognition sequence (IRS) in both the host genome and the plasmid vector to be transfected was required for this integration. The integration occurred within the IRS pre-introduced into the host genome and resulted in a complete or partial deletion of the sequence and an adjacent drug-resistant gene. This site-specific integration was not observed upon transfection without the integrase or with vectors harboring no IRS. This novel method may be useful for manipulating a mammalian genome or for targeting a retroviral genome integrated into a virus-infected cell by using the virus-specific integrase and LTR sequence.
Department of Bacterial and Blood Products, National Institute of Infectious Diseases, 1-23-1 Toyama-cho, Shinjuku-ku, Tokyo, 162-8640, Japan.
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Tanaka
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Tanaka
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Komuro
D001665:Binding Sites / Q000235:genetics; D015139:Blotting, Southern; D002460:Cell Line; D002875:Chromosomes / Q000235:genetics; D002999:Clone Cells; D004247:DNA / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D005057:Eukaryotic Cells / Q000166:cytology / Q000378:metabolism*; D016678:Genome; D019426:Integrases / Q000235:genetics*; D001909:Leukemia Virus, Bovine / Q000201:enzymology*; D010957:Plasmids / Q000235:genetics*; D011995:Recombination, Genetic; D014162:Transfection / Q000235:genetics* / Q000379:methods
D004251:DNA Transposable Elements; D004247:DNA; D019426:Integrases