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Assembly and Assessment of Prime Editing Systems for Precise Genome Editing in Plants

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CRISPR-Cas Methods

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Abstract

Prime editing, a CRISPR-Cas9-derived precise genome editing strategy, was recently developed to introduce targeted indels and all 12 types of point mutations without DNA double-strand breaks or donor DNA. The prime editing systems have been adopted for precision genome editing in crops including rice, wheat, maize, and tomato, which substantially expands the scope and capabilities of precision plant breeding. Here, we describe a fast and efficient method for construction of prime editing vectors based on Gateway assembly and efficiency assessment of prime editors through transient expression analyses in rice protoplasts.

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Acknowledgments

This work was supported by the National Science Foundation Plant Genome Research Program (award no. IOS-1758745 and IOS-2029889) and the U.S. Department of Agriculture Biotechnology Risk Assessment Grant Program (award no. 2018-33522-28789 and 2020-33522-32274).

Author information

Authors and Affiliations

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

    Simon Sretenovic, Changtian Pan & Yi** Qi

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

    Yi** Qi

Authors
  1. Simon Sretenovic
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  2. Changtian Pan
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  3. Yi** Qi
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Corresponding author

Correspondence to Yi** Qi .

Editor information

Editors and Affiliations

  1. Institute of Biotechnology and Genetic Engineering (IBGE), Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh

    M. Tofazzal Islam

  2. ICAR-National Rice Research Institute, Cuttack, Odisha, India

    Kutubuddin Ali Molla

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Sretenovic, S., Pan, C., Qi, Y. (2021). Assembly and Assessment of Prime Editing Systems for Precise Genome Editing in Plants. In: Islam, M.T., Molla, K.A. (eds) CRISPR-Cas Methods. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1657-4_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1657-4_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1656-7

  • Online ISBN: 978-1-0716-1657-4

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