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Mutagenesis in Xenopus and Zebrafish using TALENs

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TALENs

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

Abstract

Transcription activator-like effector nucleases (TALENs) have been proven to be effective for gene specific targeting across species. Here we describe the validated protocol for TALEN assembly as well as methods for generating gene knockout animals of Xenopus and zebrafish. This protocol covers selection of TALEN targeting sites, TALEN assembly with a modified Golden Gate method, injection of TALEN mRNAs into Xenopus and zebrafish embryos as well as the detection of somatic and germ-line transmitted mutations. Finally, the establishment of knockout Xenopus and zebrafish lines is also described. This protocol will facilitate broader applications of TALENs in developmental biology.

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Acknowledgments

This work is supported by grants from the Research Grants Council of Hong Kong (N_CUHK413/12 to H Z and 463013 to CHKC), the National Natural Science Foundation of China (31172394 to CHKC) and the Shenzhen Municipal R&D Fund of Science and Technology (GJHS20120702105523308 to CHKC). We thank colleagues in our laboratories for helpful discussion on this project.

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Authors and Affiliations

  1. School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China

    Yun Liu, Hui Zhao & Christopher H. K. Cheng

  2. School of Biomedical Sciences Core Laboratory, Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, 518057, China

    Yun Liu, Hui Zhao & Christopher H. K. Cheng

Authors
  1. Yun Liu
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  2. Hui Zhao
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  3. Christopher H. K. Cheng
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Corresponding authors

Correspondence to Hui Zhao or Christopher H. K. Cheng .

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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Liu, Y., Zhao, H., Cheng, C.H.K. (2016). Mutagenesis in Xenopus and Zebrafish using 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_16

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  • DOI: https://doi.org/10.1007/978-1-4939-2932-0_16

  • 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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