Transposition and gene disruption in the male germline of the mouse.
We have tested a synthetic, functional, transposon called Sleeping Beauty for use in mice as a germline insertional mutagen. We describe experiments in which mutagenic, polyadenylation-site trapping, transposon vectors were introduced into the germline of mice. When doubly transgenic males, expressing the Sleeping Beauty transposase gene (SB10) and harboring poly(A)-trap transposon vectors, were outcrossed to wild-type females, offspring were generated with new transposon insertions. The frequency of new transposon insertion is roughly two per male gamete. These new insertions can be passed through 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 insertion sites, including one insertion into a gene represented in the EST database and one in the promoter of the galactokinase (Gck) gene. This technique has promise as a new strategy for forward genetic screens in the mouse or functional genomics.
Dupuy
A J
AJ
Beckman Center for Transposon Research, University of Minnesota Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Fritz
S
S
Largaespada
D A
DA
D000818:Animals; D001483:Base Sequence; D003433:Crosses, Genetic; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005813:Genes, Synthetic; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D013094:Spermatozoa; D014170:Transformation, Genetic; D019895:Transposases / Q000235:genetics*
D004251:DNA Transposable Elements; D004247:DNA; D019895:Transposases