Open AccessGenome‐wide investigation of multiplexed CRISPR‐Cas12a‐mediated editing in rice
Author(s) -
Zhang Yingxiao,
Wu Yuechao,
Li Gen,
Qi Aileen,
Zhang Yong,
Zhang Tao,
Qi Yiping
Publication year - 2023
Publication title -
the plant genome
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.403
H-Index - 41
ISSN - 1940-3372
DOI - 10.1002/tpg2.20266
Subject(s) - crispr , genome editing , biology , genome , cas9 , palindrome , computational biology , genetics , dna sequencing , oryza sativa , genome engineering , dna , gene
Abstract 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.