title: Production of knockout rats using ENU mutagenesis and a yeast-based screening assay.
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.
authors.affiliation: Department of Oncology, McArdle Laboratory for Cancer Research, 1400 University Avenue, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
authors.forename: Yunhong
authors.initials: Y
authors.lastname: Zan
authors.forename: Jill D
authors.initials: JD
authors.lastname: Haag
authors.forename: Kai-Shun
authors.initials: KS
authors.lastname: Chen
authors.forename: Laurie A
authors.initials: LA
authors.lastname: Shepel
authors.forename: Don
authors.initials: D
authors.lastname: Wigington
authors.forename: Yu-Rong
authors.initials: YR
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Hu
authors.forename: Christine C
authors.initials: CC
authors.lastname: Lopez-Guajardo
authors.forename: Heidi L
authors.initials: HL
authors.lastname: Brose
authors.forename: Katherine I
authors.initials: KI
authors.lastname: Porter
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Leonard
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Hitt
authors.forename: Stacy L
authors.initials: SL
authors.lastname: Schommer
authors.forename: Anu F
authors.initials: AF
authors.lastname: Elegbede
authors.forename: Michael N
authors.initials: MN
authors.lastname: Gould
mesh_terms: D000818:Animals; D019313:BRCA1 Protein / Q000172:deficiency / Q000235:genetics; D024682:BRCA2 Protein / Q000172:deficiency / Q000235:genetics; D002478:Cells, Cultured; D005038:Ethylnitrosourea / Q000008:administration & dosage*; D005786:Gene Expression Regulation / Q000187:drug effects*; D018390:Gene Targeting / Q000379:methods*; D008297:Male; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering / Q000379:methods; D051381:Rats; D011922:Rats, Mutant Strains / Q000235:genetics* / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015003:Yeasts / Q000187:drug effects / Q000302:isolation & purification
chemical_list: D019313:BRCA1 Protein; D024682:BRCA2 Protein; D011994:Recombinant Proteins; D005038:Ethylnitrosourea