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

TLDR

Transgenic and gene‑targeted mutant mice are powerful tools for studying cellular processes in development and disease. The study aims to develop a site‑specific recombination strategy for generating multiple ES cell lines with tetracycline‑inducible genes at a defined locus. Using homologous recombination, an frt homing site was inserted into ES cells, enabling efficient integration of transgenes by FLPe recombinase. The approach and vectors are broadly applicable to any locus in ES cells, facilitating rapid production of mice with transgenes precisely targeted to a defined site. © 2006 Wiley‑Liss, Inc.; genesis 44:23–28, 2006.

Abstract

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.