Transposition and gene disruption in the male germline of the mouse

Adam J. Dupuy, Sabine Fritz, David A. Largaespada

Research output: Contribution to journalArticle

177 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)82-88
Number of pages7
JournalGenesis
Volume30
Issue number2
DOIs
StatePublished - Jul 16 2001

Keywords

  • Insertional mutagenesis
  • Sleeping Beauty
  • Transposon

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