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.
Publication types
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Evaluation Study
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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BRCA1 Protein / deficiency
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BRCA1 Protein / genetics
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BRCA2 Protein / deficiency
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BRCA2 Protein / genetics
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Cells, Cultured
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Ethylnitrosourea / administration & dosage*
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Gene Expression Regulation / drug effects*
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Gene Targeting / methods*
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Male
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Mutagenesis, Site-Directed*
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Protein Engineering / methods
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Rats
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Rats, Mutant Strains / genetics*
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Rats, Mutant Strains / metabolism
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Rats, Sprague-Dawley
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Yeasts / drug effects
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Yeasts / isolation & purification
Substances
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BRCA1 Protein
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BRCA2 Protein
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Recombinant Proteins
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Ethylnitrosourea