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Inhibition of human immunodeficiency virus type 1 by RNA interference using long-hairpin RNA

Abstract

Inhibition of virus replication by means of RNA interference has been reported for several important human pathogens, including human immunodeficiency virus type 1 (HIV-1). RNA interference against these pathogens has been accomplished by introduction of virus-specific synthetic small interfering RNAs (siRNAs) or DNA constructs encoding short-hairpin RNAs (shRNAs). Their use as therapeutic antiviral against HIV-1 is limited, because of the emergence of viral escape mutants. In order to solve this durability problem, we tested DNA constructs encoding virus-specific long-hairpin RNAs (lhRNAs) for their ability to inhibit HIV-1 production. Expression of lhRNAs in mammalian cells may result in the synthesis of many siRNAs targeting different viral sequences, thus providing more potent inhibition and reducing the chance of viral escape. The lhRNA constructs were compared with in vitro diced double-stranded RNA and a DNA construct encoding an effective nef-specific shRNA for their ability to inhibit HIV-1 production in cells. Our results show that DNA constructs encoding virus-specific lhRNAs are capable of inhibiting HIV-1 production in a sequence-specific manner, without inducing the class I interferon genes.

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Acknowledgements

RNAi research in the Berkhout lab is sponsored by NWO-CW (TOP grant) and Senter-NOVEM (TS grant with Viruvation). We thank Stephan Heynen for performing CA-p24 ELISA; Dr P Midoux (Centre de Biophysique Moleculaire, Orleans, France) for providing us with the pT7-luc plasmid; Dr Jean-Marc Jacque (University of Massachusetts Medical School, USA) for providing the pcDNA3-T7pol expressing vector and Dr E de Jong (University of Amsterdam, The Netherlands) for the kind gift of IFN-β primers.

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Correspondence to B Berkhout.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Konstantinova, P., de Vries, W., Haasnoot, J. et al. Inhibition of human immunodeficiency virus type 1 by RNA interference using long-hairpin RNA. Gene Ther 13, 1403–1413 (2006). https://doi.org/10.1038/sj.gt.3302786

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