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Plant Genome Editing with CRISPR Systems pp 245–256Cite as

Plant Gene Knockout and Knockdown by CRISPR-Cpf1 (Cas12a) Systems

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

Abstract

CRISPR-Cpf1 (Cas12a) is a class II type V endonuclease, which has been used as a genome editing tool in different biological systems. Here we describe a fast, efficient, and user-friendly system for CRISPR-Cpf1 expression vector assembly. In this system, the Pol II promoter is used to drive the expression of both Cpf1 and its crRNA, with the crRNA flanked by hammerhead (HH) and hepatitis delta virus (HDV) ribozyme RNAs for precise crRNA processing. All the components of this system can be modified depending on plant species and experimental goals. Using this system, nearly 100% editing efficiency and 90% gene expression decrease were achieved in rice and Arabidopsis, respectively.

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Acknowledgements

This work is supported by funds from University of Maryland and Syngenta Biotechnology.

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Authors and Affiliations

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

    Yingxiao Zhang & Yiping Qi

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

    Yong Zhang

  3. Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD, USA

    Yiping Qi

Authors
  1. Yingxiao Zhang
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  2. Yong Zhang
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Corresponding author

Correspondence to Yiping Qi .

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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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Zhang, Y., Zhang, Y., Qi, Y. (2019). Plant Gene Knockout and Knockdown by CRISPR-Cpf1 (Cas12a) Systems. 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_18

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  • DOI: https://doi.org/10.1007/978-1-4939-8991-1_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8990-4

  • Online ISBN: 978-1-4939-8991-1

  • eBook Packages: Springer Protocols

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