Production of knockout rats using ENU mutagenesis and a yeast-based screening assay |
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| Authors: | Zan Yunhong Haag Jill D Chen Kai-Shun Shepel Laurie A Wigington Don Wang Yu-Rong Hu Rong Lopez-Guajardo Christine C Brose Heidi L Porter Katherine I Leonard Rachel A Hitt Andrew A Schommer Stacy L Elegbede Anu F Gould Michael N |
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| Institution: | Department of Oncology, McArdle Laboratory for Cancer Research, 1400 University Avenue, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA. |
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| Abstract: | 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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