Negative feedback inhibition of HIV-1 by TAT-inducible expression of siRNA.
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.
Unwalla
Hoshang J
HJ
Li
Ming-Jie
MJ
Kim
James D
JD
Li
Hai Tang
HT
Ehsani
Ali
A
Alluin
Jessica
J
Rossi
John J
JJ
D002478:Cells, Cultured; D005246:Feedback / Q000502:physiology; D015696:Gene Products, tat / Q000096:biosynthesis* / Q000235:genetics*; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods; D015497:HIV-1 / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000378:metabolism / Q000821:virology; D009000:Monocytes / Q000378:metabolism / Q000821:virology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism / Q000821:virology*; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D018840:HSP70 Heat-Shock Proteins; D034741:RNA, Small Interfering; D054322:tat Gene Products, Human Immunodeficiency Virus