Nucleic Acids Research Advance Access originally published online on August 31, 2006
Nucleic Acids Research 2006 34(16):4527-4536; doi:10.1093/nar/gkl628
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Nucleic Acids Research, 2006, Vol. 34, No. 16 4527-4536
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Molecular Biology |
Tumor inhibition by genomically integrated inducible RNAi-cassettes
Department of Gynecology and Obstetrics, School of Medicine, J.W. Goethe-University Theodor-Stern-Kai 7, D-60590 Frankfurt, Germany
*To whom correspondence should be addressed. Tel: +49 69 6301 6894; Fax: +49 69 6301 6364; Email: Strebhardt{at}em.uni-frankfurt.de
Received July 21, 2006. Accepted August 10, 2006.
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.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  D. J. Adams and L. van der Weyden Contemporary approaches for modifying the mouse genome Physiol Genomics, August 1, 2008; 34(3): 225 - 238. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Henriksen, C. Lokke, M. Hammero, D. Geerts, R. Versteeg, T. Flaegstad, and C. Einvik Comparison of RNAi efficiency mediated by tetracycline-responsive H1 and U6 promoter variants in mammalian cell lines Nucleic Acids Res., May 14, 2007; 35(9): e67 - e67. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Seibler, A. Kleinridders, B. Kuter-Luks, S. Niehaves, J. C. Bruning, and F. Schwenk Reversible gene knockdown in mice using a tight, inducible shRNA expression system Nucleic Acids Res., April 1, 2007; 35(7): e54 - e54. [Abstract] [Full Text] [PDF]
|
|