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Hitz, C. ; Steuber-Buchberger, P.M. ; Delic, S. ; Wurst, W. ; Kühn, R.

Generation of shRNA transgenic mice.

Methods Mol. Biol. 530, 101-129 (2009)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
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
Language
Publication Year 2009
HGF-reported in Year 2009
ISSN (print) / ISBN 1064-3745
e-ISSN 1940-6029
ISBN 978-1-934115-26-8
Journal Methods in Molecular Biology
Quellenangaben Volume: 530, Issue: , Pages: 101-129 Article Number: , Supplement: ,
Series Methods Mol. Biol.
Publisher Springer
Publishing Place Berlin [u.a.]
Reviewing status Peer reviewed
Institute(s) Institute of Developmental Genetics (IDG)
POF-Topic(s) 30204 - Cell Programming and Repair
Research field(s) Genetics and Epidemiology
PSP Element(s) G-500500-001
DOI 10.1007/978-1-59745-471-1_6
Scopus ID 65549123686
PubMed ID 19266327
Erfassungsdatum 2009-09-21
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