Conventional approaches to produce transgenic mice recurrently yield unpredictable patterns and levels of transgene expression, a situation calling for the development of new techniques to overcome these drawbacks in the context of overexpression studies. Here we present an efficient method for rap
Conditional transgene expression mediated by the mouse β-actin locus
✍ Scribed by Ulrike Jägle; Jürg A. Gasser; Matthias Müller; Bernd Kinzel
- Book ID
- 102155342
- Publisher
- John Wiley and Sons
- Year
- 2007
- Tongue
- English
- Weight
- 408 KB
- Volume
- 45
- Category
- Article
- ISSN
- 1526-954X
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
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 β‐actin locus. Flp/FRT 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 β‐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. © 2007 Wiley‐Liss, Inc.
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