Published online 13 February 2004
Nucleic Acids Research, 2004, Vol. 32, No. 3 1154-1158
© 2004 Oxford University Press
Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins
Laboratory of Retrovirology, Division of Infectious Diseases, Department of Medicine, Brown Medical School, 4th Floor, 55 Claverick Street, Providence, RI 02903, USA
*To whom correspondence should be addressed. Tel: +1 401 444 5219; Fax: +1 401 444 2939; Email: bramratnam{at}lifespa.org
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Post-transcriptional inhibition of HIV-1 replication can be achieved by RNA interference (RNAi). The cellular expression of short interfering RNA (siRNA) or short hairpin RNA (shRNA) homologous to regions of the HIV-1 genome decreases viral replication by the selective degradation of targeted RNA. Here, we demonstrate that another class of noncoding regulatory RNA, termed microRNA (miRNA), can be used to deliver antiviral RNAi. By incorporating sequences encoding siRNA targeting the HIV-1 transactivator protein tat into a human miR-30 pre-microRNA (pre-miRNA) backbone, we were able to express tat siRNA in cells. The tat siRNA delivered as pre-miRNA precursor was 80% more effective in reducing HIV-1 p24 antigen production than tat siRNA expressed as conventional shRNA. Our results confirm the utility of expressing HIV-1 specific siRNA through a miR-30 precursor stem–loop structure and suggest that this strategy can be used to increase the antiviral potency of RNAi.
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