Generation of a stable cell line producing high-titer self-inactivating lentiviral vectors.
To facilitate the generation of SIN lentivirus vector-producer cell lines, we have developed a novel conditional SIN (cSIN) lentivirus vector, which retains its SIN properties in normal target cells yet can be produced at high titers from tetracycline-regulated packaging cell lines. The design of the cSIN vector is based on replacing the vector U3 transcription regulatory elements with the Tet-responsive element, which allows vector production exclusively in cells expressing the synthetic Tet-regulated transactivator (tTA). In contrast minimal vector production ( approximately 200 IU/ml) is obtained in target cells that do not express the tTA, even in the presence of all HIV-1 proteins. Following transduction of the Tet-regulated SODk1 lentivirus vector-packaging cell line with the cSIN vector, high titers of cSIN recombinant vector (>10(6) IU/ml) could be generated, which efficiently transduced terminally differentiated neurons in normal rat brain.
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Gene Therapy Center, University of North Carolina Chapel Hill, 7119 Thurston-Bowles, Chapel Hill, North Carolina 27599-7352, USA.
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McCown
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Verma
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Kafri
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D000818:Animals; D002460:Cell Line; D005434:Flow Cytometry; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000378:metabolism; D008954:Models, Biological; D008957:Models, Genetic; D009474:Neurons / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats; D014158:Transcription, Genetic; D014161:Transduction, Genetic; D014162:Transfection
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins