Site-specific recombinases provide a means of efficiently manipulating
chromosomal sequences in mammalian cells in culture and in mice.
Embryonic stem cells containing recombinase nucleic acid constructs that
were expressed in the male germline would simplify current protocols for
producing mice bearing homologously recombined alleles that have been
secondarily rearranged by a site-specific recombinase, five lines of
transgenic mice containing a fusion gene consisting of the mouse
protamine 1 gene promoter and the Cre recombinase coding sequence (ProCre
nucleic acid constructs) showed high levels of Cre-mediated recombination
of the germline, but did not show appreciable recombination in other
tissues. In different ProCre strains, between 80% and 100% of the progeny
that inherited a Cre target nucleic acid construct from males that were
also heterozygous for a ProCre nucleic acid construct inherited the
Cre-recombined target. ProCre nucleic acid constructs and recombined
targets segregated in the first generation. When ES cells prepared from
one ProCre line were transfected with vectors containing a loxP-flanked
neomycin cassette, G418 resistant, homologously recombined clones, in
which the loxP sites remained functional, were readily isolated. These
data establish that ProCre nucleic acid constructs will facilitate the
production of subtle, conditional, or tissue-specific mutations in mice
as well as the production and analysis of mice with
recombinase-conditional lethal alleles.