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Epigenome Editing pp 89–109Cite as

Generation of TALE-Based Designer Epigenome Modifiers

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1767))

Abstract

Manipulation of gene expression can be facilitated by editing the genome or the epigenome. Precise genome editing is traditionally achieved by using designer nucleases which are generally exploited to eliminate a specific gene product. Upon the introduction of a site-specific DNA double-strand break (DSB) by the nuclease, endogenous DSB repair mechanisms are in turn harnessed to induce DNA sequence changes that can result in target gene inactivation. Minimal off-target effects can be obtained by endowing designer nucleases with the highly specific DNA-binding domain (DBD) derived from transcription activator-like effectors (TALEs). In contrast, epigenome editing allows gene expression control without inducing changes in the DNA sequence by specifically altering epigenetic marks, as histone tails modifications or DNA methylation patterns within promoter or enhancer regions. Importantly, this approach allows both up- and downregulation of the target gene expression, and the effect is generally reversible. TALE-based designer epigenome modifiers combine the high specificity of TALE-derived DBDs with the power of epigenetic modifier domains to induce fast and long-lasting changes in the epigenetic landscape of a target gene and control its expression. Here we provide a detailed description for the generation of TALE-based designer epigenome modifiers and of a suitable reporter cell line to easily monitor their activity.

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Acknowledgments

We thank the members of our laboratory for helpful discussion. This work was supported by the Federal Ministry of Education and Research (BMBF).

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Authors and Affiliations

  1. Institute for Cell and Gene Therapy & Center for Chronic Immunodeficiency, Medical Center—University of Freiburg, Freiburg, Germany

    Sandra Nitsch & Claudio Mussolino

Authors
  1. Sandra Nitsch
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  2. Claudio Mussolino
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Corresponding author

Correspondence to Claudio Mussolino .

Editor information

Editors and Affiliations

  1. Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, Stuttgart, Germany

    Albert Jeltsch

  2. Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands

    Marianne G. Rots

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Nitsch, S., Mussolino, C. (2018). Generation of TALE-Based Designer Epigenome Modifiers. In: Jeltsch, A., Rots, M. (eds) Epigenome Editing. Methods in Molecular Biology, vol 1767. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7774-1_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7774-1_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7773-4

  • Online ISBN: 978-1-4939-7774-1

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