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Negative feedback inhibition of HIV-1 by TAT-inducible expression of siRNA

Nature Biotechnology volume 22, pages 15731578 (2004) | Download Citation

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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.

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Acknowledgements

This research was supported by National Institutes of Health grants AI 29329, AI42552 and HL074704.

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Affiliations

  1. Division of Molecular Biology, Beckman Research Institute of The City of Hope, Duarte, California 91010, USA.

    • Hoshang J Unwalla
    • , Ming-Jie Li
    • , James D Kim
    • , Hai Tang Li
    • , Ali Ehsani
    • , Jessica Alluin
    •  & John J Rossi

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The authors declare no competing financial interests.

Corresponding author

Correspondence to John J Rossi.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Transcription occurs from both, HIV-1 LTR and mhsp70 TATA boxes.

  2. 2.

    Supplementary Fig. 2

    Partial processing of shRNA loop.

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DOI

https://doi.org/10.1038/nbt1040

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