Production of knockout rats using ENU mutagenesis and a yeast-based screening assay

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 deve...

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Bibliographic Details
Published in:	Nature biotechnology Vol. 21; no. 6; pp. 645 - 651
Main Authors:	Gould, Michael N, 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
Format:	Journal Article
Language:	English
Published:	New York, NY Nature 01-06-2003 Nature Publishing Group
Subjects:	Animals Biological and medical sciences Biotechnology BRCA1 Protein - deficiency BRCA1 Protein - genetics BRCA2 Protein - deficiency BRCA2 Protein - genetics Breast cancer Cells, Cultured Ethylnitrosourea - administration & dosage Fundamental and applied biological sciences. Psychology Gene Expression Regulation - drug effects Gene Targeting - methods Male Mutagenesis, Site-Directed Mutation Physiology Protein Engineering - methods Rats Rats, Mutant Strains - genetics Rats, Mutant Strains - metabolism Rats, Sprague-Dawley Recombinant Proteins - genetics Recombinant Proteins - metabolism Rodents Yeasts Yeasts - drug effects Yeasts - isolation & purification Screening Vertebrata Mammalia Yeast Mutagenesis Rat Rodentia Production
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Summary:	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.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	1087-0156 1546-1696
DOI:	10.1038/nbt830