ZFIN ID: ZDB-PUB-090422-21
Highly Efficient ENU Mutagenesis in Zebrafish
de Bruijn, E., Cuppen, E., and Feitsma, H.
Date: 2009
Source: Methods in molecular biology (Clifton, N.J.)   546: 3-12 (Chapter)
Registered Authors: Cuppen, Edwin, de Bruijn, Ewart, Feitsma, Harma
Keywords: Zebrafish, ENU, Mutagenesis, TILLING, Method, Mutant, Knockout
MeSH Terms:
  • Alkylating Agents/administration & dosage
  • Animals
  • Animals, Genetically Modified
  • DNA/drug effects
  • Drug Administration Schedule
  • Ethylnitrosourea/administration & dosage*
  • Female
  • Genetic Engineering/methods
  • Genome/drug effects
  • Germ-Line Mutation
  • Male
  • Mutagenesis/drug effects*
  • Zebrafish/genetics*
PubMed: 19378094 Full text @ Meth. Mol. Biol.
ENU (N-ethyl-N-nitrosourea) mutagenesis is a widely accepted and proven method to introduce random point mutations in the genome. Because there are no targeted knockout strategies available for zebrafish so far, random mutagenesis is currently the preferred method in both forward and reverse genetic approaches. To obtain high-density mutagenized zebrafish, six consecutive ENU treatments are applied at weekly intervals to adult male zebrafish by bathing them in ENU solution. With this procedure an average germ line mutation load of one mutation every 1.0 x 10(5)-1.5 x 10(5) basepairs is reached routinely in our lab.