Production of knockout rats using ENU mutagenesis and a yeast-based screening assay. Nat Biotechnol 2003 Jun;21(6):645-51
Date
05/20/2003Pubmed ID
12754522DOI
10.1038/nbt830Scopus ID
2-s2.0-0037685273 (requires institutional sign-in at Scopus site) 190 CitationsAbstract
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.
Author List
Zan Y, Haag JD, Chen KS, Shepel LA, Wigington D, Wang YR, Hu R, Lopez-Guajardo CC, Brose HL, Porter KI, Leonard RA, Hitt AA, Schommer SL, Elegbede AF, Gould MNAuthor
Anuoluwapo F. Elegbede MD Assistant Professor in the Surgery department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBRCA1 Protein
BRCA2 Protein
Cells, Cultured
Ethylnitrosourea
Gene Expression Regulation
Gene Targeting
Male
Mutagenesis, Site-Directed
Protein Engineering
Rats
Rats, Mutant Strains
Rats, Sprague-Dawley
Recombinant Proteins
Yeasts
