Gene silencing using micro-RNA designed hairpins

Michael T. McManus, Christian P. Petersen, Brian B. Haines, Jianzhu Chen, Phillip A. Sharp*

*Corresponding author for this work

Research output: Contribution to journalArticle

246 Scopus citations

Abstract

During RNA interference (RNAi), long dsRNA is processed to ∼21 nt duplexes, short interfering RNAs (siRNAs), which silence genes through a mRNA degradation pathway. Small temporal RNAs (stRNAs) and micro-RNAs (miRNAs) are ∼21 nt RNAs that are processed from endogenously encoded hairpin-structured precursors, and function to silence genes via translational repression. Here we report that synthetic hairpin RNAs that mimic siRNAs and miRNA precursor molecules can target a gene for silencing, and the mechanism of silencing appears to be through mRNA degradation and not translational repression. The sequence and structural configuration of these RNAs are important, and even slight modification in structure can affect the silencing activity of the hairpins. Furthermore, these RNAs are active when expressed by DNA vectors containing polymerase III promoters, opening the possibility for new approaches in stable RNAi-based loss of function studies.

Original languageEnglish (US)
Pages (from-to)842-850
Number of pages9
JournalRNA
Volume8
Issue number6
DOIs
StatePublished - Jun 22 2002

Keywords

  • H1 promoter
  • RNAi
  • T-cell
  • dsRNA
  • miRNA
  • siRNA

ASJC Scopus subject areas

  • Molecular Biology

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    McManus, M. T., Petersen, C. P., Haines, B. B., Chen, J., & Sharp, P. A. (2002). Gene silencing using micro-RNA designed hairpins. RNA, 8(6), 842-850. https://doi.org/10.1017/S1355838202024032