International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


ORAL PRESENTATION

MONDAY OCTOBER 18

4.45pm – 5.00pm

REGION-SPECIFIC SATURATED MUTAGENESIS USING THE SLEEPING BEAUTY TRANSPOSON SYSTEM

Takeda J1, Keng VW1, Yae K1, Hayakawa T1, Mizuno S1, Uno Y2, Kokubu C1, Horie K1

1 Dept. of Social and Environmental Medicine, Osaka University Graduate School of Medicine, Osaka, Japan, 2 The Institute of Experimental Animal Science, Osaka University, Osaka, Japan

Generation of mutant mice is important in determining the function of genes in the post-genome era. Sleeping Beauty (SB) is a synthetic transposon derived from a fish genome and we have previously demonstrated that the SB transposon sytem worked efficiently in mouse germ line, suggesting a potential usage of a large-scale mutagenesis in mice. We also observed that about 80% of transposition events in germ cell preferentially occurred near the donor site and within the same chromosome.  After screening about 600 mice, the number of transposon-integrated gene hits was shown to be clustered within approximately 3 Mb distance of the donor site.  From a double-positive (containing SB transposase and transposon, acting as a mutagen) transgenic mouse with donor site located on chromosome 12, we obtained 16 clustered transposon integrations, corresponding to about 50% of known genes located within the 3 Mb region.  This indicates the usefulness of this system for saturated mutagenesis, which is currently impossible with other large-scale mutagenic systems. We are currently analyzing about 50 mutant lines with transposon integration in both known and novel genes by generating homozygotes and analyzing the resulting phenotype. Taken together, SB transposon system is a rapid and efficient method for region-specific mutagenesis.

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