International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


POSTER 124 - DYNAMIC INSERTIONAL MUTAGENESIS PROJECT FOR THE MOUSE

Poirier C, Overbeek PA, Adams CP, Harrison WR, Xiao N, Castile CA, Bishop CE

Baylor College of Medicine, Houston, United States

We developed a 2-step insertional mutagenesis program for the mouse using the FVB/N albino inbred strain. All transgenes carried a visible semidominant coat color marker, tyrosinase, flanked by the inverted terminal repeats, specific for the Sleeping Beauty transposase. In the first step, we generated regular transgenic mice with our tyrosinase transgenes and transmission of the transgene was followed with the visible coat color marker. In the second step, the transgene was transposed from its original insertion site through the “cut and paste” system mediated by the Sleeping Beauty transposase. For transpositional purposes, we generated different transgenic lines in the FVB/N background which expressed the transposase from 4 different promotors. Transposition in the paternal germline was identified by new variations of pigmentation in the progeny. In both steps, heterozygous and homozygous mice were easily identified by their coat color and were screened for inherited abnormal phenotypes. From this phenotype driven mutagenesis program, the causative genes were subsequently cloned using the transgene/transposon as a probe. Several tyrosinase transgene constructs have been generated. The lastest versions contain a LoxP site for further engineering, inversion and deletion within the mouse genome and 2 splice acceptors in opposite orientation for in vivo gene trapping. We will discuss and compare the transposition and mutagenesis efficiency of all the transgenes.

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