TY - JOUR
T1 - Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector
AU - van de Wetering, Marc
AU - Oving, Irma
AU - Muncan, Vanesa
AU - Pon Fong, Menno Tjon
AU - Brantjes, Helen
AU - van Leenen, Dik
AU - Holstege, Frank C.P.
AU - Brummelkamp, Thijn R.
AU - Agami, Reuven
AU - Clevers, Hans
PY - 2003/6/1
Y1 - 2003/6/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=10744219596&partnerID=8YFLogxK
U2 - 10.1038/sj.embor.embor865
DO - 10.1038/sj.embor.embor865
M3 - Article
C2 - 12776180
AN - SCOPUS:10744219596
SN - 1469-221X
VL - 4
SP - 609
EP - 615
JO - EMBO Reports
JF - EMBO Reports
IS - 6
ER -