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

Marc van de Wetering, Irma Oving, Vanesa Muncan, Menno Tjon Pon Fong, Helen Brantjes, Dik van Leenen, Frank C.P. Holstege, Thijn R. Brummelkamp, Reuven Agami, Hans Clevers

Onderzoeksoutput: Bijdrage aan tijdschriftArtikelpeer review

465 Citaten (Scopus)

Samenvatting

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 β-catenin in colorectal cancer (CRC) cells. T-cell factor (TCF) target-gene expression is induced by accumulated β-catenin, and is the main transforming event in these cells. We have shown previously that the disruption of β-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 β-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.

Originele taal-2Engels
Pagina's (van-tot)609-615
Aantal pagina's7
TijdschriftEMBO Reports
Volume4
Nummer van het tijdschrift6
DOI's
StatusGepubliceerd - 1 jun. 2003
Extern gepubliceerdJa

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