Nucleic Acids Research Advance Access originally published online on March 21, 2007
Nucleic Acids Research 2007 35(7):e54; doi:10.1093/nar/gkm122
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Nucleic Acids Research, 2007, Vol. 35, No. 7 e54
© 2007 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.
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Reversible gene knockdown in mice using a tight, inducible shRNA expression system
1Artemis Pharmaceuticals GmbH, Neurather Ring 1, 51063 Cologne, Germany, 2Institute for Genetics, Department of Mouse Genetics and Metabolism and Center for Molecular Medicine (CMMC), University of Cologne, Germany and 3University of Applied Science Gelsenkirchen, Department of Applied Natural Sciences, August-Schmidt-Ring 10, 45665 Recklinghausen, Germany
*To whom correspondence should be addressed. Tel: +49-221-9645342; Fax: +49-221-9645321; Email: j.seibler{at}artemispharma.de
Received October 30, 2006. Revised February 13, 2007. Accepted February 13, 2007.
RNA interference through expression of short hairpin (sh)RNAs provides an efficient approach for gene function analysis in mouse genetics. Techniques allowing to control time and degree of gene silencing in vivo, however, are still lacking. Here we provide a generally applicable system for the temporal control of ubiquitous shRNA expression in mice. Depending on the dose of the inductor doxycycline, the knockdown efficiency reaches up to 90%. To demonstrate the feasibility of our tool, a mouse model of reversible insulin resistance was generated by expression of an insulin receptor (Insr)-specific shRNA. Upon induction, mice develop severe hyperglycemia within seven days. The onset and progression of the disease correlates with the concentration of doxycycline, and the phenotype returns to baseline shortly after withdrawal of the inductor. On a broad basis, this approach will enable new insights into gene function and molecular disease mechanisms.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
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