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

Nucleic Acids Res. 2003 Sep 1;31(17):5033-8. doi: 10.1093/nar/gkg704.

Abstract

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 tRNA(met)-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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line
  • Gene Expression Regulation, Viral
  • Gene Products, tat / genetics
  • HIV-1 / genetics*
  • Humans
  • Promoter Regions, Genetic / genetics*
  • RNA Interference*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • RNA, Transfer, Met / genetics
  • RNA, Viral / genetics
  • RNA, Viral / metabolism
  • Transfection
  • tat Gene Products, Human Immunodeficiency Virus

Substances

  • Gene Products, tat
  • RNA, Small Interfering
  • RNA, Transfer, Met
  • RNA, Viral
  • tat Gene Products, Human Immunodeficiency Virus