Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells.
The production of subtle or conditional mutations in mice through the combined use of site-specific and homologous recombination has become an increasingly widespread experimental paradigm in mammalian genetics. Embryonic stem cells containing recombinase transgenes that were expressed in the male germ line, but not in other tissues or in the embryonic stem cells themselves, would substantially simplify the production of such alleles. Here we show that transgenes comprised of the mouse protamine 1 promoter and the Cre recombinase coding sequence mediate the efficient recombination of a Cre target transgene in the male germ line, but not in other tissues. Embryonic stem cell lines generated from one of these transgenic strains were transfected with targeting vectors that included loxP-flanked selectable markers, and homologously recombined alleles containing the marker and functional loxP sites were isolated. These results establish the potential of the system for substantially reducing the time, effort, and resources required to produce homologously recombined alleles in mice that have been secondarily rearranged by a site-specific recombinase.
O'Gorman
S
S
Gene Expression Laboratory, The Salk Institute for Biological Studies, San Diego, CA 92186, USA. ogorman@salk.edu
Dagenais
N A
NA
Qian
M
M
Marchuk
Y
Y
D000483:Alleles; D000818:Animals; D015870:Gene Expression; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D005854:Germ Cells; D019426:Integrases / Q000235:genetics*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011479:Protamines / Q000235:genetics*; D011995:Recombination, Genetic; D013234:Stem Cells; D014764:Viral Proteins
C513176:Prm1 protein, mouse; D011479:Protamines; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases