RNA



Gene silencing using micro-RNA designed hairpins


MICHAEL T.  McMANUS  a1, CHRISTIAN P.  PETERSEN  a1, BRIAN B.  HAINES  a1, JIANZHU  CHEN  a1 and PHILLIP A.  SHARP  a1 c1
a1 Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA

Abstract

During RNA interference (RNAi), long dsRNA is processed to [similar]21 nt duplexes, short interfering RNAs (siRNAs), which silence genes through a mRNA degradation pathway. Small temporal RNAs (stRNAs) and micro-RNAs (miRNAs) are [similar]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.

(Received March 25 2002)
(Revised April 3 2002)
(Accepted April 9 2002)


Key Words: dsRNA; H1 promoter; miRNA; RNAi; siRNA; T-cell.

Correspondence:
c1 Reprint requests to: Phillip A. Sharp, Center for Cancer Research, Massachusetts Institute of Technology, 40 Ames Street E17-526, Cambridge, Massachusetts 02139, USA; e-mail: sharppa@mit.edu.