Intron-Based Single Transcript Unit CRISPR Systems for Plant Genome Editing

Zhaohui Zhong; Shishi Liu; Xiaopei Liu; Binglin Liu; Xu Tang; Qiurong Ren; Jianping Zhou; Xuelian Zheng; Yiping Qi; Yong Zhang

doi:10.1186/s12284-020-0369-8

Rice (Feb 2020)

Intron-Based Single Transcript Unit CRISPR Systems for Plant Genome Editing

Zhaohui Zhong,
Shishi Liu,
Xiaopei Liu,
Binglin Liu,
Xu Tang,
Qiurong Ren,
Jianping Zhou,
Xuelian Zheng,
Yiping Qi,
Yong Zhang

Affiliations

Zhaohui Zhong: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Shishi Liu: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Xiaopei Liu: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Binglin Liu: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Xu Tang: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Qiurong Ren: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Jianping Zhou: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Xuelian Zheng: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China
Yiping Qi: Department of Plant Science and Landscape Architecture, University of Maryland
Yong Zhang: Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China

DOI: https://doi.org/10.1186/s12284-020-0369-8
Journal volume & issue: Vol. 13, no. 1
pp. 1 – 13

Abstract

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Abstract Background Expression of either Cas9 or Cas12a and guide RNAs by a single Polymerase II (Pol II) promoter represents a compact CRISPR expression system and has many advantages for different applications. In order to make this system routine in plant biology, engineering efforts are needed for developing and optimizing such single transcript unit (STU) systems for plant genome editing. Results To develop novel intron-based STU (iSTU) CRISPR system (STU CRISPR 3.0), we first evaluated three introns from three plant species for carrying guide RNAs by using an enhanced green fluorescence protein (eGFP) system in rice. After validation of proper intron slicing, we inserted these gRNA-containing introns into the open reading frames (ORFs) of Cas9 and Cas12a for testing their genome editing capability. Different guide RNA processing strategies have been tested for Cas9 and Cas12a. We demonstrated singular genome editing and multiplexed genome editing with these iSTU-Cas9 and iSTU-Cas12a systems. Conclusion We developed multiple iSTU-CRISPR/Cas9 and Cas12a systems for plant genome editing. Our results shed light on potential directions for further improvement of the iSTU systems.

Published in Rice

ISSN: 1939-8425 (Print); 1939-8433 (Online)
Publisher: SpringerOpen
Country of publisher: United Kingdom
LCC subjects: Agriculture: Plant culture
Website: https://thericejournal.springeropen.com/

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