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J. Virol., 09 1997, 7128-7131, Vol 71, No. 9
Copyright © 1997, American Society for Microbiology

A conditional self-inactivating retrovirus vector that uses a tetracycline-responsive expression system

JJ Hwang, L Li and WF Anderson
Gene Therapy Laboratories, Norris Cancer Center, University of Southern California School of Medicine, Los Angeles 90033, USA.

We developed a novel conditional self-inactivating (C-SIN) vector, TL- SN, by replacement of the enhancer-promoter of the 3' long terminal repeat of Moloney murine leukemia virus with a synthetic tetracycline operator-cytomegalovirus promoter (tetP) from the tetracycline- responsive expression system (TRES). The other component of the TRES, a chimeric transactivator (tTA), was stably incorporated into PA317 amphotropic packaging cells, thus generating the packaging cell line PA317-tTA. C-SIN amphotropic G418-resistant virus particles were generated with a titer of 2 x 10(5) CFU/ml within 2 days of transinfection of PA317-tTA cells with TL-SN ecotropic virus particles. This titer was approximately 2 log units higher than that obtained by transinfection of parental PA317 cells and was due to the high level of viral transcripts originating from the tetP promoter at the 5' end of the transduced vector in the presence of tTA. Our C-SIN vector has the potential for use in human gene therapy since it incorporates the advantages of previous SIN vectors in having weak tetP promoter activity (in the absence of tTA in target cells) while at the same time achieving high viral titers with PA317-tTA packaging cells.

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