PUBLICATION

Target-selected inactivation of the zebrafish rag1 gene

Authors
Wienholds, E., Schulte-Merker, S., Walderich, B., and Plasterk, R.H.A.
ID
ZDB-PUB-020716-5
Date
2002
Source
Science (New York, N.Y.)   297(5578): 99-102 (Journal)
Registered Authors
Plasterk, Ronald H.A., Schulte-Merker, Stefan, Wienholds, Erno
Keywords
none
MeSH Terms
  • Amino Acid Substitution
  • Animals
  • Codon, Terminator
  • Ethylnitrosourea
  • Female
  • Gene Library
  • Gene Rearrangement
  • Genes, Immunoglobulin
  • Genes, RAG-1*
  • Haplotypes
  • Heterozygote
  • Homeodomain Proteins/chemistry
  • Homeodomain Proteins/genetics
  • Immunoglobulin Heavy Chains/genetics
  • Introns
  • Male
  • Mutagenesis
  • Mutation*
  • Mutation, Missense
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide
  • Recombination, Genetic
  • Zebrafish/genetics*
  • Zebrafish/immunology
  • Zebrafish/physiology
PubMed
12098699 Full text @ Science
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 F(1) 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 F(1) fish were collected and cryopreserved, we have in principle generated a mutant library from which mutants of most zebrafish genes can be isolated.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes