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Biotech / Medical : RNAi -- Ignore unavailable to you. Want to Upgrade?

To: nigel bates who wrote (30)	6/3/2003 1:13:05 PM
From: tuck	Read Replies (1) \| Respond to of 671

>>chopping off the tails An unfortunate choice of words.<< Three SKID mice, three SKID mice, See their RNAP II. Their own promoter took their life, They cut off its tails with a DICER knife. Their expression is no longer rife. Three SKID mice . . . Sorry, must have been the coffee. Here's another method using a tweaked RNA polymerase promoter as the vector, but this time RNA polymerase III instead of II. No mention of how they tweaked it in the abstract. >>EMBO Rep 2003 Jun;4(6):609-15 Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector. Van De Wetering M, Oving I, Muncan V, Pon Fong MT, Brantjes H, Van Leenen D, Holstege FC, Brummelkamp TR, Agami R, Clevers H. [1] Hubrecht Laboratory, Centre for Biomedical Genetics, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands [2] These authors contributed equally to this work. 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.<< Cheers, Tuck