Nature Biotechnology21, 645 - 651 (2003)
Published online: 18 May 2003; | doi:10.1038/nbt830
Production of knockout rats using ENU mutagenesis and a yeast-based
screening assay
Yunhong Zan1, 2, Jill D Haag1, 2, Kai-Shun Chen1, Laurie A Shepel1, Don Wigington1, Yu-Rong Wang1, Rong Hu1, Christine C Lopez-Guajardo1, Heidi L Brose1, Katherine I Porter1, Rachel A Leonard1, Andrew A Hitt1, Stacy L Schommer1, Anu F Elegbede1
& Michael N Gould1
1
Department of Oncology, McArdle Laboratory for
Cancer Research, 1400 University Avenue, University of
Wisconsin−Madison, Madison, Wisconsin
53706, USA.
The rat is a widely used model in biomedical research and is often
the preferred rodent model in many areas of physiological and pathobiological
research. Although many genetic tools are available for the rat, methods to
produce gene-disrupted knockout rats are greatly needed. In this study, we
developed protocols for creating N-ethyl-N-nitrosourea
(ENU)-induced germline mutations in several rat strains. F1
preweanling pups from mutagenized Sprague Dawley (SD) male rats were then
screened for functional mutations in Brca1 and Brca2 using a
yeast gap-repair, ADE2-reporter truncation assay. We produced knockout
rats for each of these two breast cancer suppressor genes.
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