A toolkit for transgenesis at the ROSA26 locus by recombinase-mediated cassette exchange

Abstract

We describe a toolkit to perform transgenesis at the ROSA26 locus by recombinase-mediated cassette exchange that will eliminate the inherent problem of random insertion via traditional pronuclear injection.  A recombination site-tagged embryonic stem (ES) cell line and two cloning vectors were constructed to facilitate the generation of targeted ES cells with the transgene of interest at ROSA26.  The experimental procedure is simple and efficient, and can be readily adapted in many laboratories for rapid generation of transgenic mice.