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- Title
Target-selected inactivation of the zebrafish rag1 gene.
- Authors
Wienholds, Erno; Schulte-Merker, Stefan; Walderich, Brigitte; Plasterk, Ronald H A
- Abstract
The zebrafish has become a favorite organism for genetic analysis of vertebrate development, but methods for generating mutants by reverse genetic approaches have been lacking. We report a method to obtain stable mutants of a gene based on knowledge of the gene sequence only. Parental fish were mutagenized with N-ethyl-N-nitrosourea; in 2679 F1 fish, the rag1 gene was analyzed for heterozygous mutations by resequencing. In total, we found 15 mutations: 9 resulted in amino acid substitutions and 1 resulted in a premature stop codon. This truncation mutant was found to be homozygous viable and defective in V(D)J joining. Although presumably immune deficient, these homozygous rag1 mutant fish are able to reach adulthood and are fertile. As sperm samples from all 2679 F1 fish were collected and cryopreserved, we have in principle generated a mutant library from which mutants of most zebrafish genes can be isolated.
- Publication
Science (New York, N.Y.), 2002, Vol 297, Issue 5578, p99
- ISSN
1095-9203
- Publication type
Journal Article
- DOI
10.1126/science.1071762