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Enzyme Engineering pp 63–72Cite as

TAL Effector Nuclease (TALEN) Engineering

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 978))

Abstract

TALENs, fusion proteins of DNA binding domains of TAL (transcription activator-like) effectors and the DNA cleavage domains of endonuclease FokI, have emerged as genetic tools for targeted gene modification, holding great potential for basic and applied research, even for gene therapy. Here we present a simple and efficient approach to custom-engineering TALEN genes with four basic TAL repeats and their DNA recognition cipher. The “modular assembly” method also involves the “Golden Gate” cloning strategy, using 53 ready-to-use plasmids in just two rounds of restriction and ligation to assemble TALENs with up to 24 repeat units that recognize up to 24 bp of target DNA.

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Acknowledgment

This work was supported by US National Science Foundation Plant Genome Research Project (award no. 0820831 to BY).

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Authors and Affiliations

  1. Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, USA

    Ting Li & Bing Yang

Authors
  1. Ting Li
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  2. Bing Yang
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Corresponding author

Correspondence to Bing Yang .

Editor information

Editors and Affiliations

  1. New England Biolabs, Inc., County Road 240, Ipswich, 01938, Massachusetts, USA

    James C. Samuelson

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Li, T., Yang, B. (2013). TAL Effector Nuclease (TALEN) Engineering. In: Samuelson, J. (eds) Enzyme Engineering. Methods in Molecular Biology, vol 978. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-293-3_5

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  • DOI: https://doi.org/10.1007/978-1-62703-293-3_5

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-292-6

  • Online ISBN: 978-1-62703-293-3

  • eBook Packages: Springer Protocols

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