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Nucleic Acids Research Advance Access originally published online on June 22, 2007
Nucleic Acids Research 2007 35(12):e90; doi:10.1093/nar/gkm475
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Nucleic Acids Research, 2007, Vol. 35, No. 12 e90
© 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.

Methods Online

Conditional brain-specific knockdown of MAPK using Cre/loxP regulated RNA interference

Christiane Hitz1, Wolfgang Wurst1,2 and Ralf Kühn1,*

1GSF National Research Center for Environment and Health, Institute of Developmental Genetics, Ingolstaedter Landstrasse 1, 85764 Neuherberg and 2Max-Planck-Institute of Psychiatry, Molecular Neurogenetics, Kraepelinstrasse 2-10, 80804 Munich, Germany

*To whom correspondence should be addressed. Tel: +49 89 3187 2887; Fax: +49 89 3187 3099; Email: ralf.kuehn{at}gsf.de

Received April 18, 2007. Revised May 30, 2007. Accepted May 30, 2007.

In the last years, RNA interference (RNAi)-mediated gene knockdown has developed into a routine method to assess gene function in cultured mammalian cells in a fast and easy manner. For the use of this technique in developing or adult mice, short hairpin (sh)RNA vectors expressed stably from the genome are a faster alternative to conventional knockout approaches. Here we describe an advanced strategy for conditional gene knockdown in mice, where we used the Cre/loxP system to activate RNAi in a time and tissue dependent manner in the adult mouse brain. By placing conditional RNAi constructs into the defined genomic Rosa26 locus and by using recombinase mediated cassette exchange (RMCE) instead of laborious homologous recombination, we developed a fast, easy and reproducible approach to assess gene function in adult mice. We applied this technique to three genes of the MAPK signaling pathway—Braf, Mek1 and Mek2—and demonstrate here the potential of this new tool in mouse mutagenesis.


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