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Nucleic Acids Research, 2003, Vol. 31, No. 17 5033-5038
© 2003 Oxford University Press

Promoter choice affects the potency of HIV-1 specific RNA interference

Daniel Boden, Oliver Pusch, Fredrick Lee, Lynne Tucker, Peter R. Shank and Bharat Ramratnam*

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}Lifespan.org
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

RNA interference (RNAi) is mediated by small interfering (si) RNAs that target and degrade mRNA in a sequence-specific manner. Cellular expression of siRNA can be achieved by the use of expression cassettes driven by RNA polymerase III (pol III) promoters. Here, we demonstrate that a modified tRNAmet-derived (MTD) promoter effectively drives the cellular expression of HIV-1-specific siRNA. We observed up to 56% greater inhibition of virus production when the MTD promoter was used to drive the expression of short hairpin (sh) RNA targeting the HIV-1 transactivator protein tat compared to cassettes containing other pol III promoters such as H1, U6+1 and U6+27. We conclude that the MTD promoter is ideally suited to drive intracellular expression of HIV-1 specific siRNA and may serve as an important component of future RNAi vector delivery systems.


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