SpringerProtocols: Abstract: Highly Efficient ENU Mutagenesis in Zebrafish

Highly Efficient ENU Mutagenesis in Zebrafish

By: Ewart de Bruijn⁴, Edwin Cuppen⁴, Harma Feitsma⁴

Abstract

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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.

Affiliation(s): (4) Hubrecht Institute for Developmental Biology and Stem Cell Research, Uppsalalaan 8, 3584, CT, Utrecht, The Netherlands

Book Title: Zebrafish: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 546 | Pub. Date: Jan-01-2010 | Page Range: 3-12 | DOI: 10.1007/978-1-60327-977-2_1

Subject: Biochemistry

Key Words: Zebrafish - ENU - Mutagenesis - TILLING - Method - Mutant - Knockout

References

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