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Zebrafish

Volume 546 of the series Methods in Molecular Biology pp 3-12

Date:

Highly Efficient ENU Mutagenesis in Zebrafish

  • Ewart de BruijnAffiliated withHubrecht Institute for Developmental Biology and Stem Cell Research
  • , Edwin CuppenAffiliated withHubrecht Institute for Developmental Biology and Stem Cell Research
  • , Harma FeitsmaAffiliated withHubrecht Institute for Developmental Biology and Stem Cell Research

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Summary

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 × 105–1.5 × 105 basepairs is reached routinely in our lab.

Key words

Zebrafish ENU Mutagenesis TILLING Method Mutant Knockout