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Design, Construction, and Validation of Targeted Gene Activation with TREE System in Human Cells

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Epigenomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2577))

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Abstract

Genome editing technologies can be diverted into artificial transcription activators. In particular, researchers have improved dCas9-based technologies by tandem-fusing or trans-accumulating effector domains. Previously, we developed a hierarchical effector accumulation system named “TREE,” enabling robust activation of target genes even when strongly silenced. In this chapter, we describe our protocol to design, construct, and validate the TREE-mediated target gene activation in cultured human cells.

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Acknowledgments

We thank Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript. This work was funded by Grant-in-Aid for JSPS Fellows (grant# 20J13292) to A.K. and Grant-in-Aid for Early-Career Scientists (grant# 19K16111) to T.S.

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Authors and Affiliations

  1. Division of Integrated Sciences for Life, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan

    Atsushi Kunii, Takashi Yamamoto & Tetsushi Sakuma

Authors
  1. Atsushi Kunii
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  2. Takashi Yamamoto
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  3. Tetsushi Sakuma
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Corresponding author

Correspondence to Tetsushi Sakuma .

Editor information

Editors and Affiliations

  1. Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan

    Izuho Hatada

  2. Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan

    Takuro Horii

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Kunii, A., Yamamoto, T., Sakuma, T. (2023). Design, Construction, and Validation of Targeted Gene Activation with TREE System in Human Cells. In: Hatada, I., Horii, T. (eds) Epigenomics. Methods in Molecular Biology, vol 2577. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2724-2_15

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  • DOI: https://doi.org/10.1007/978-1-0716-2724-2_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2723-5

  • Online ISBN: 978-1-0716-2724-2

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