International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


POSTER 92 - INDUCIBLE GENE TRAPPING USING DRUG SELECTABLE MARKERS AND THE CRE/LOXP SYSTEM TO IDENTIFY DEVELOPMENTALLY REGULATED GENES

YT Chen
Baylor College of Medicine

1) Liu P, 1) Vaishishnav S, 2) Bradley A
1) Baylor College of Medicine, 2) The Welcome Trust Sanger Institute

Gene trapping in mouse ES cells is an important functional genomics approach. It allows identification of novel genes and the generation of corresponding mutant mice at the same time. To develop a system which allows in vitro prescreening of gene traps that respond to signaling molecules of interest, we designed a selection scheme using drug selectable markers and the Cre/loxP technology to perform in vitro prescreening in a 96-well format. In our experiment, 920 gene trap clones were assayed, and 258 of them were classified as gene traps induced by in vitro differentiation. Sixty-five of the in vitro differentiation inducible gene traps were also responsive to retinoic acid (RA) treatment. An analysis of 13 RA-inducible gene trap clones in vivo reveals that nearly 85% (11 out of 13) of the RA-inducible gene traps trapped developmentally regulated genes. This result is consistent with the observation that genes induced by RA treatment are likely to be developmentally regulated(citationInduction gene trap). Our experiment demonstrates that the inducible gene trapping system we established can be used to enrich for gene traps of interest in vitro.


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