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.
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Acknowledgments
The authors would like to thank Freek van Eeden for details provided on the mutagenesis protocol. This work was supported by funds from the European Union-funded FP6 Integrated Project ZF-MODELS.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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de Bruijn, E., Cuppen, E., Feitsma, H. (2009). Highly Efficient ENU Mutagenesis in Zebrafish. In: Lieschke, G., Oates, A., Kawakami, K. (eds) Zebrafish. Methods in Molecular Biology, vol 546. Humana Press. https://doi.org/10.1007/978-1-60327-977-2_1
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DOI: https://doi.org/10.1007/978-1-60327-977-2_1
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