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A Rapid TALEN Assembly Protocol

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Polycomb Group Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1480))

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Abstract

Owing to their modular and highly specific DNA recognition mode, transcription activator-like effector nucleases (TALENs) have been rapidly adopted by the scientific community for the purpose of generating site-specific double-strand breaks (DSBs) on a DNA molecule. A pair of TALENs can be used to produce random insertions or deletions of various lengths via nonhomologous end-joining or together with a homologous donor DNA to induce precise sequence alterations by homologous recombination (HR). Here, we describe a method for TALEN assembly (easyT) and a strategy for genome engineering via HR.

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Acknowledgments

We thank Makiko Seimiya for help with embryo microinjections. Work of T.K. and R.P. was supported by a grant from the Deutsche Forschungsgemeinschaft SPP1356—“Pluripotency and Cellular Reprogramming.” Work in the R.P. laboratory is supported by ETH Zürich.

Author information

Authors and Affiliations

  1. Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule Zürich, Mattenstrasse 26, 4058, Basel, Switzerland

    Arslan Akmammedov & Renato Paro

  2. Department of Genetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1 Bunkyo-ku, 113-8654, Tokyo, Japan

    Tomonori Katsuyama

  3. Faculty of Science, University of Basel, Klingelbergstrasse 50, 4056, Basel, Switzerland

    Renato Paro

Authors
  1. Arslan Akmammedov
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  2. Tomonori Katsuyama
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  3. Renato Paro
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Corresponding author

Correspondence to Arslan Akmammedov .

Editor information

Editors and Affiliations

  1. IBCN-CNR, Institute of Cell Biology and Neurobiology and Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, Rome, Italy

    Chiara Lanzuolo

  2. Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, Milan, Italy

    Beatrice Bodega

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Akmammedov, A., Katsuyama, T., Paro, R. (2016). A Rapid TALEN Assembly Protocol. In: Lanzuolo, C., Bodega, B. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 1480. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6380-5_23

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  • DOI: https://doi.org/10.1007/978-1-4939-6380-5_23

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6378-2

  • Online ISBN: 978-1-4939-6380-5

  • eBook Packages: Springer Protocols

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