Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector.

Abstract:

:We have designed a doxycycline-regulated form of the H1 promoter of RNA polymerase III that allows the inducible knockdown of gene expression by small interfering RNAs (siRNAs). As a proof-of-principle, we have targeted beta-catenin in colorectal cancer (CRC) cells. T-cell factor (TCF) target-gene expression is induced by accumulated beta-catenin, and is the main transforming event in these cells. We have shown previously that the disruption of beta-catenin/TCF4 activity in CRC cells by the overexpression of dominant-negative TCF induces rapid G1 arrest and differentiation. Stable integration of our inducible siRNA vector allowed the rapid production of siRNAs on doxycycline induction, followed by specific downregulation of beta-catenin. In these CRC cells, TCF reporter-gene activity was inhibited, and G1 arrest and differentiation occurred. The inhibition of two other genes using this vector system shows that it should be useful for the inducible knockdown of gene expression.

journal_name

EMBO Rep

journal_title

EMBO reports

authors

van de Wetering M,Oving I,Muncan V,Pon Fong MT,Brantjes H,van Leenen D,Holstege FC,Brummelkamp TR,Agami R,Clevers H

doi

10.1038/sj.embor.embor865

keywords:

subject

Has Abstract

pub_date

2003-06-01 00:00:00

pages

609-15

issue

6

eissn

1469-221X

issn

1469-3178

pii

embor865

journal_volume

4

pub_type

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