Efficient method to generate single‐copy transgenic mice by site‐specific integration in embryonic stem cells

C Beard, K Hochedlinger, K Plath, A Wutz… - genesis, 2006 - Wiley Online Library
C Beard, K Hochedlinger, K Plath, A Wutz, R Jaenisch
genesis, 2006Wiley Online Library
Transgenic and gene‐targeted mutant mice provide powerful tools for analysis of the cellular
processes involved in early development and in the pathogenesis of many diseases. Here
we describe a transgene integration strategy mediated by site‐specific recombination that
allows establishment of multiple embryonic stem (ES) cell lines carrying tetracycline‐
inducible genes targeted to a specific locus to assure predictable temporal and spatial
expression in ES cells and mice. Using homologous recombination we inserted an frt …
Abstract
Transgenic and gene‐targeted mutant mice provide powerful tools for analysis of the cellular processes involved in early development and in the pathogenesis of many diseases. Here we describe a transgene integration strategy mediated by site‐specific recombination that allows establishment of multiple embryonic stem (ES) cell lines carrying tetracycline‐inducible genes targeted to a specific locus to assure predictable temporal and spatial expression in ES cells and mice. Using homologous recombination we inserted an frt homing site into which tetracycline‐inducible transgenes can be integrated efficiently in the presence of FLPe recombinase. This strategy and the vectors described here are generally applicable to any locus in ES cells and should allow for the rapid production of mice with transgenes efficiently targeted to a defined site. genesis 44:23–28, 2006. © 2006 Wiley‐Liss, Inc.
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