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
Citation
Wienholds, E., Schulte-Merker, S., Walderich, B., and Plasterk, R.H.A. (2002) Target-selected inactivation of the zebrafish rag1 gene. Science (New York, N.Y.). 297(5578):99-102.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping