Abstract
Zebrafish is a valuable model organism to study vertebrate development, organ regeneration and to generate human disease models. As an important member of the arsenal of genome editing, TALE nucleases (TALENs) have implicated in broad applications in zebrafish reverse genetic studies. In this chapter, we describe the detailed protocols of TALEN-mediated genome manipulations in zebrafish, including targeted gene disruption by indel mutations, deletion of large genomic regions by using two pairs of TALENs, and precise genome modification by homologous recombination (HR).
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Acknowledgments
We thank Yan Shen and Yuying Gao for the management of TALEN constructs; Yingdi Jia and **gliang Chen for maintenance of our zebrafish facility; and all the members in our lab for their efforts on optimizing the TALEN applications. This work was partially supported by the National Natural Science Foundation of China (NSFC) (31110103904) and the National Key Basic Research Program of China (2012CB945101, 2011CBA01000, 2015CB942803). This work was also supported by the NSFC grant 81371264.
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Huang, P., **ao, A., Tong, X., Lin, S., Zhang, B. (2016). Targeted Mutagenesis in Zebrafish by TALENs. In: Kühn, R., Wurst, W., Wefers, B. (eds) TALENs. Methods in Molecular Biology, vol 1338. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2932-0_15
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DOI: https://doi.org/10.1007/978-1-4939-2932-0_15
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2931-3
Online ISBN: 978-1-4939-2932-0
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