Journal
GENESIS
Volume 45, Issue 11, Pages 659-666Publisher
WILEY
DOI: 10.1002/dvg.20342
Keywords
RMCE; targeted transgenesis; mouse; beta-actin; conditional expression
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Transgenic mice are an effective model to study gene function in vivo; however, position effects can complicate tissue-specific transgene analysis. To facilitate precise targeting of a transgenic construct into the mouse genome, we combined the Cre/lox and Flp/ FRT recombination systems to allow for rapid transgene replacement and conditional transgene expression from the endogenous P-actin locus. FIp/FIRT recombination was used to rapidly exchange FRT-flanked transgene cassettes by recombinase-mediated cassette exchange in embryonic stem cells, while transgene expression can be activated in mice after Cre-mediated excision of a floxed STOP cassette. To validate our system, we analyzed the expression profile of an EGFP reporter gene after integration into the P-actin locus and Cre-mediated excision of the floxed STOP cassette. Breeding of EGFP reporter, mice with various Cre mouse lines resulted in the expected expression profiles, demonstrating the feasibility of the model to facilitate predictable and strong transgene expression in a spatially and temporally controlled manner. genesis 45:659-666, 2007. (c) 2007 Wiley-Liss, Inc.
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