Generation of rat mutants using a coat color-tagged <Emphasis Type="Italic">Sleeping Beauty</Emphasis> transposon system |
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| Authors: | Baisong Lu Aron M Geurts Christophe Poirier Deborah C Petit Wilbur Harrison Paul A Overbeek Colin E Bishop |
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| Institution: | (1) Department of Obstetrics and Gynecology, Baylor College of Medicine, Houston, Texas 77030, USA;(2) Institute for Regenerative Medicine, Wake Forest University Health Sciences, Winston-Salem, North Carolina 27157, USA;(3) Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA;(4) Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA;(5) Present address: Department of Pediatrics, University of Chicago, Chicago, IL 60637, USA;(6) Present address: Department of Veterinary Medicine and Surgery, University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030, USA |
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| Abstract: | A significant barrier to exploiting the full potential of the rat as a biomedical model is the lack of tools to easily modify
its germline. Here we show that a tyrosinase-tagged Sleeping Beauty transposon can be used as a simple, efficient method to generate rat mutants in vivo. By making two lines of transgenic rats, one carrying the transposon and another expressing the transposase in germ cells,
we are able to obtain bigenic males in which transposition occurs in the germ cells. We show that transposition leads to the
appearance of new coat colors in the offspring. Using such bigenic males, we obtained an average of 1.2 transpositions per
gamete and identified 19 intragenic integration events among 96 transposition sites that were sequenced. In addition, gene
trapping was confirmed and rats with evidence for transposon-induced dominant ocular anomalies were identified. These data
suggest that the modified Sleeping Beauty transposon represents a powerful new tool for producing molecularly defined mutagenesis in the rat. |
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