Genome-wide investigation of multiplexed CRISPR-Cas12a-mediated editing in rice.

Autor: Zhang Y; Dep. of Plant Science and Landscape Architecture, Univ. of Maryland, College Park, MD, 20742, USA., Wu Y; Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Jiangsu Key Laboratory of Crop Genetics and Physiology, Agricultural College of Yangzhou Univ., Yangzhou, 225009, China.; Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou, 225009, China., Li G; Dep. of Plant Science and Landscape Architecture, Univ. of Maryland, College Park, MD, 20742, USA., Qi A; Dep. of Plant Science and Landscape Architecture, Univ. of Maryland, College Park, MD, 20742, USA., Zhang Y; Dep. of Biotechnology, School of Life Science and Technology, Center for Informational Biology, Univ. of Electronic Science and Technology of China, Chengdu, 610054, China., Zhang T; Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Jiangsu Key Laboratory of Crop Genetics and Physiology, Agricultural College of Yangzhou Univ., Yangzhou, 225009, China.; Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou, 225009, China.; Key Laboratory of Plant Functional Genomics of the Ministry of Education/Joint International Research Laboratory of Agriculture and Agri-Product Safety, The Ministry of Education of China, Yangzhou Univ., Yangzhou, 225009, China., Qi Y; Dep. of Plant Science and Landscape Architecture, Univ. of Maryland, College Park, MD, 20742, USA.; Institute for Bioscience and Biotechnology Research, Univ. of Maryland, Rockville, MD, 20850, USA.
Jazyk: angličtina
Zdroj: The plant genome [Plant Genome] 2023 Jun; Vol. 16 (2), pp. e20266. Date of Electronic Publication: 2022 Sep 30.
DOI: 10.1002/tpg2.20266
Abstrakt: Clustered regularly interspaced short palindromic repeats (CRISPR) nucleases like Cas9 and Cas12a are revolutionizing plant basic research and crop breeding. A major advantage of CRISPR over earlier nucleases systems is its capability of multiplexed genome editing. However, it remains unknown about the potential off-target effects when multiple concurrent DNA double-strand breaks (DSBs) are induced in a crop genome. Here, we investigated this important question in rice (Oryza sativa) using a highly multiplexed CRISPR-Cas12a system. With whole-genome sequencing, we first revealed high genome editing specificity of Mb2Cas12a and protospacer adjacent motif promiscuity of LbCas12a. We discovered large chromosomal rearrangement events in edited rice plants that endured many (e.g., >50) simultaneous DSBs, but not in plants that endured lower order DSBs (e.g., <10). Our results shed important light on the analysis and regulation of engineered crops derived from CRISPR-Cas mediated multiplexed genome editing.
(© 2022 The Authors. The Plant Genome published by Wiley Periodicals LLC on behalf of Crop Science Society of America.)
Databáze: MEDLINE
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