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Gene silencing using micro-RNA designed hairpins

Published online by Cambridge University Press:  20 August 2002

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

Keywords

dsRNAH1 promotermiRNARNAisiRNAT-cell
Type
Research Article
Information
RNA , Volume 8 , Issue 6 , June 2002 , pp. 842 - 850
Copyright
© 2002 RNA Society

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