Transposition and gene disruption in the male germline of the mouse

We have tested a synthetic, functional, transposon called Sleeping Beauty f or use in mice as a germline insertional mutagen, We describe experiments i n which mutagenic, polyadenylation-site trapping, transposon vectors were i ntroduced into the germline of mice. When doubly transgenic males, expressi ng the Sleeping Beauty transposase gene (SB10) and harboring poly(A)-trap t ransposon vectors, were outcrossed to wild-type females, offspring were gen erated with new transposon insertions. The frequency of new transposon inse rtion is roughly two per male gamete. These new insertions can be passed th rough the germline to the next generation and can insert into or near genes . We have generated a preliminary library of 24 mice harboring 56 novel ins ertion sites, including one insertion into a gene represented in the EST da tabase and one in the promoter of the galactokinase (Gck) gene. This techni que has promise as a new strategy for forward genetic screens in the mouse or functional genomics, (C) 2001 Wiley-Liss, Inc.