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Plant Genome Editing with CRISPR Systems pp 75–82Cite as

A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1917))

Abstract

The CRISPR-Cas9 system has been widely adopted in genome editing. By changing the 20 bp guide sequence, it can easily edit any sequence adjacent to a protospacer adjacent motif (PAM) in a genome. Multiplex genome editing could be accomplished with simultaneous expression of multiple single-guide RNAs (sgRNA). Given that sgRNAs are expressed by Pol III promoters, multiplex genome editing is conventionally done by assembly of multiple complete sgRNA expression cassettes together, which can be a challenge in vector construction. Here, we described a multiplex genome editing system based on a single transcript unit CRISPR-Cas9 (STU CRISPR-Cas9) expression system driven by a single Pol II promoter. It represents a novel approach for multiplex genome editing.

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Acknowledgments

This work was supported by grants to YZ from the Sichuan Youth Science and Technology Foundation (2017JQ0005), the National Science Foundation of China (31771486), the National Transgenic Major Project (2018ZX08022001-003), and the Fundamental Research Funds for the Central Universities (ZYGX2016J119 and ZYGX2016J122).

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Authors and Affiliations

  1. Department of Biotechnology, School of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Chengdu, China

    Xu Tang, Zhaohui Zhong, Qiurong Ren, Binglin Liu & Yong Zhang

Authors
  1. Xu Tang
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  2. Zhaohui Zhong
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  3. Qiurong Ren
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  4. Binglin Liu
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  5. Yong Zhang
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Corresponding author

Correspondence to Yong Zhang .

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Editors and Affiliations

  1. Department of Plant Science and Landscape Architecture, University of Maryland College Park, College Park, MD, USA

    Yiping Qi

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Tang, X., Zhong, Z., Ren, Q., Liu, B., Zhang, Y. (2019). A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants. In: Qi, Y. (eds) Plant Genome Editing with CRISPR Systems. Methods in Molecular Biology, vol 1917. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8991-1_6

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  • DOI: https://doi.org/10.1007/978-1-4939-8991-1_6

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8990-4

  • Online ISBN: 978-1-4939-8991-1

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