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Targeted Mutagenesis in Zebrafish by TALENs

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TALENs

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1338))

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 Jingliang 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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Authors and Affiliations

  1. Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, 100871, P. R. China

    Peng Huang, An Xiao, Xiangjun Tong & Bo Zhang

  2. Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA, 90095, USA

    Shuo Lin

  3. College of Life Sciences, Peking University, No 5 YiHeYuan Rd., Haidian District, Beijing, 100871, P. R. China

    Bo Zhang

Authors
  1. Peng Huang
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  2. An Xiao
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  3. Xiangjun Tong
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  4. Shuo Lin
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  5. Bo Zhang
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Corresponding author

Correspondence to Bo Zhang .

Editor information

Editors and Affiliations

  1. Max-Delbrück-Center for Molecular Medicine, Berlin, Germany

    Ralf Kühn

  2. Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany

    Wolfgang Wurst

  3. Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Germany

    Benedikt Wefers

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Huang, P., Xiao, 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

  • eBook Packages: Springer Protocols

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