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RNA. 2005 Feb;11(2):220-6. Epub 2004 Dec 21.

Overexpression of exportin 5 enhances RNA interference mediated by short hairpin RNAs and microRNAs.

Author information

  • 1Department of Molecular Genetics and Microbiology, Duke University Medical Center, Box 3025, Room 426 CARL Building, Research Drive, Durham, NC 27710, USA.

Abstract

Plasmids or viral vectors that express short hairpin RNAs (shRNAs) have emerged as important tools for the stable inhibition of specific genes by RNA interference. shRNAs are structural and functional homologs of pre-microRNAs, intermediates in the production of endogenously encoded microRNAs (miRNAs). Therefore, overexpressed shRNAs could inhibit miRNA function by competing for a limiting level of one or more factors involved in miRNA biogenesis or function. Here, we demonstrate that overexpressed shRNAs can saturate the activity of endogenous Exportin 5, a factor required for nuclear export of both shRNAs and pre-miRNAs. While shRNA overexpression can therefore inhibit miRNA function, simultaneous overexpression of Exportin 5 reverses this effect. Moreover, Exportin 5 overexpression can significantly enhance RNA interference mediated by shRNAs. These data have implications for the future clinical utilization of shRNAs and also provide a simple method to enhance RNA interference by shRNAs in culture.

PMID:
15613540
[PubMed - indexed for MEDLINE]
PMCID:
PMC1370710
Free PMC Article

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