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Delivery of Designer Epigenome Modifiers into Primary Human T Cells

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Epigenome Editing

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

Abstract

The development of tools which allow for the precise alterations of the epigenetic landscape in desired genomic locations presents exciting possibilities toward further understanding how gene expression is regulated and opportunities to harness these properties for therapeutic purposes. In contrast to gene knockout strategies, targeted epigenome modifications, such as editing of DNA methylation, can mediate gene expression modulation without changing the genomic sequence. Thereby, in a therapeutic context, this strategy may offer a safer route as compared to gene disruption using designer nucleases that, to reach high efficiencies, relies on the occurrence of random mutations to inactivate the target gene. In addition, therapeutic benefit is influenced not only by the intrinsic safety and efficacy of the tools used but also by methods that allow efficient and non-toxic transfer of the selected reagents in the target cells. Here, we describe a detailed protocol, for safe delivery of TALE-based designer epigenome modifiers in the form of in vitro transcribed mRNA into primary human CD4+ T cells to efficiently silence the expression of an exemplary human gene (i.e., CCR5).

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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) and the German Academic Exchange Service (DAAD).

Author information

Authors and Affiliations

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

    Tafadzwa Mlambo, Marianna Romito, Tatjana I. Cornu & Claudio Mussolino

  2. Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, Germany

    Tafadzwa Mlambo

Authors
  1. Tafadzwa Mlambo
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  2. Marianna Romito
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  3. Tatjana I. Cornu
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  4. 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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Mlambo, T., Romito, M., Cornu, T.I., Mussolino, C. (2018). Delivery of Designer Epigenome Modifiers into Primary Human T Cells. 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_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7774-1_9

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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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