Latest documents http://publikationen.ub.uni-frankfurt.de/index/index/ Sat, 11 Mar 2006 16:45:09 +0100 Sat, 11 Mar 2006 16:45:09 +0100 http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/2161 RNA interference (RNAi) has emerged as a powerful tool to induce loss-of-function phenotypes by post-transcriptional silencing of gene expression. In this study we wondered whether inducible RNAi-cassettes integrated into cellular DNA possess the power to trigger neoplastic growth. For this purpose inducible RNAi vectors containing tetracycline (Tet)-responsive derivatives of the H1 promoter for the conditional expression of short hairpin RNA (shRNA) were used to target human polo-like kinase 1 (Plk1), which is overexpressed in a broad spectrum of human tumors. In the absence of doxycycline (Dox) HeLa clones expressing TetR, that carry the RNAi-cassette stably integrated, exhibited no significant alteration in Plk1 expression levels. In contrast, exposure to Dox led to marked downregulation of Plk1 mRNA to 3% and Plk1 protein to 14% in cell culture compared to mismatch shRNA/Plk1-expressing cells. As a result of Plk1 depletion cell proliferation decreased to 17%. Furthermore, for harnessing RNAi for silencing disease-related genes in vivo we transplanted inducible RNAi-HeLa cells onto nude mice. After administration of Dox knockdown of Plk1 expression was observed correlating to a significant inhibition of tumor growth. Taken together, our data revealed that genomically integrated RNAi-elements are suitable to hamper tumor growth by conditional expression of shRNA. Sven Kappel; Yves Matthess; Brigitte Zimmer; Manfred Kaufmann; Klaus Strebhardt article http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/2161 Fri, 03 Nov 2006 16:45:09 +0100