1. Negative feedback inhibition of HIV-1 by TAT-inducible expression of siRNA.
- Author
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Unwalla HJ, Li MJ, Kim JD, Li HT, Ehsani A, Alluin J, and Rossi JJ
- Subjects
- Cells, Cultured, Feedback physiology, Gene Silencing, Genetic Therapy methods, HSP70 Heat-Shock Proteins genetics, Humans, Kidney metabolism, Kidney virology, Monocytes metabolism, Promoter Regions, Genetic genetics, T-Lymphocytes metabolism, tat Gene Products, Human Immunodeficiency Virus, Gene Products, tat biosynthesis, Gene Products, tat genetics, HIV-1 physiology, HSP70 Heat-Shock Proteins metabolism, Monocytes virology, RNA, Small Interfering genetics, T-Lymphocytes virology
- Abstract
Here we demonstrate that an inducible anti-HIV short hairpin RNA (shRNA) expressed from a Pol II promoter inhibits HIV-1 gene expression in mammalian cells. Our strategy is based on a promoter system in which the HIV-1 LTR is fused to the Drosophila hsp70 minimal heat shock promoter. This system is inducible by HIV-1 TAT, which functions in a negative feedback loop to activate transcription of an shRNA directed against HIV-1 rev. Upon induction the shRNA is processed to an siRNA that guides inhibition of HIV replication in cultured T-lymphocytes and hematopoietic stem cell-derived monocytes. The fusion promoter system may be safer than drug-inducible systems for shRNA-mediated gene therapy against HIV as the shRNAs are only expressed following HIV infection.
- Published
- 2004
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