Generation of a Stable Cell Line Producing High-Titer Self-Inactivating Lentiviral Vectors
2001; Elsevier BV; Volume: 3; Issue: 1 Linguagem: Inglês
10.1006/mthe.2000.0238
ISSN1525-0024
AutoresKai-lin Xu, Hong Ma, Thomas J. McCown, Inder M. Verma, Tal Kafri,
Tópico(s)CRISPR and Genetic Engineering
ResumoTo 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 (∼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 (>106 IU/ml) could be generated, which efficiently transduced terminally differentiated neurons in normal rat brain.
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