Nucleic Acids Research Advance Access published online on June 22, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm475
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Methods Online |
Conditional brain-specific knockdown of MAPK using Cre/loxP regulated RNA interference
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.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  F. P.L. Lai, M. Szczodrak, J. M. Oelkers, M. Ladwein, F. Acconcia, S. Benesch, S. Auinger, J. Faix, J. V. Small, S. Polo, et al. Cortactin Promotes Migration and Platelet-derived Growth Factor-induced Actin Reorganization by Signaling to Rho-GTPases Mol. Biol. Cell, July 15, 2009; 20(14): 3209 - 3223. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Nyabi, M. Naessens, K. Haigh, A. Gembarska, S. Goossens, M. Maetens, S. De Clercq, B. Drogat, L. Haenebalcke, S. Bartunkova, et al. Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells Nucleic Acids Res., April 1, 2009; 37(7): e55 - e55. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Burtscher and H. Lickert Foxa2 regulates polarity and epithelialization in the endoderm germ layer of the mouse embryo Development, March 15, 2009; 136(6): 1029 - 1038. [Abstract] [Full Text] [PDF]
|
|