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Open Access Green as soon as Postprint is submitted to ZB.
Generation of shRNA transgenic mice.
Methods Mol. Biol. 530, 101-129 (2009)
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 RNAi in mice, short hairpin (sh) RNAs expressed stably from the genome are a faster alternative to conventional knockout approaches. Here, we describe an advanced strategy for complete or conditional gene knockdown in mice, where the Cre/loxP system is used to activate RNAi in a time- and tissue-dependent manner. Single-copy RNAi constructs are placed into the Rosa26 locus of ES cells by recombinase-mediated cassette exchange and transmitted through the germline of chimaeric mice. The shRNA transgenic offspring can be either directly used for phenotypic analysis or are further crossed to a Cre transgenic strain to activate conditional shRNA vectors. The site-specific insertion of single-copy shRNA vectors allows the expedite and reproducible production of knockdown mice and provides an easy and fast approach to assess gene function in vivo.
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Publication type
Article: Journal article
Document type
Scientific Article
Editors
Kühn, R.* ; Wurst, W.*
Keywords
RNAi; transgenic mice; Rosa26; Cre/loxP; RMCE; shRNA
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
ISBN
978-1-934115-26-8
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 530,
Pages: 101-129
Series
Methods Mol. Biol.
Publisher
Springer
Publishing Place
Berlin [u.a.]
Reviewing status
Peer reviewed
Institute(s)
Institute of Developmental Genetics (IDG)