Efficient Induction of Point Mutations Allowing Recovery of Specific Locus Mutations in Zebrafish

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 92; no. 13; pp. 5997 - 6001
• Main Authors: Riley, Bruce B., Grunwald, David J.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 20.06.1995; National Acad Sciences; National Academy of Sciences
• Subjects: Alleles; Animals; Crosses, Genetic; Danio rerio; Dose-Response Relationship, Drug; Embryo, Nonmammalian - physiology; Embryonic and Fetal Development - drug effects; Embryonic and Fetal Development - genetics; Embryos; Ethylnitrosourea - toxicity; Female; Fish; Freshwater; Genetic loci; Genetic mutation; Genetic screening; Genetics; Germ cells; Male; Mosaic; Mutagenesis; Mutation; Pigmentation; Point Mutation; Skin Pigmentation - genetics; Time Factors; Zebrafish - embryology; Zebrafish - genetics
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.13.5997

A technique is described that greatly increases the efficiency of recovering specific locus point mutations in zebrafish (Danio rerio). Founder individuals that were mosaic for point mutations were produced by mutagenizing postmeiotic gametes with the alkylating agent N-ethyl-N-nitrosourea. Under optimal conditions, each founder carried an average of 10 mutations affecting genes required for embryogenesis. Moreover, ≈2% of these founders transmitted new mutations at any prespecified pigmentation locus. Analyses of new pigmentation mutations confirmed that most were likely to be point mutations. Thus, mutagenesis of postmeiotic gametes with N-ethyl-N-nitrosourea yielded frequencies of point mutations at specific loci that were 10- to 15-fold higher than previously achieved in zebrafish. Our procedure should, therefore, greatly facilitate recovery of multiple mutant alleles at any locus of interest.