期刊
MOLECULAR PLANT PATHOLOGY
卷 23, 期 4, 页码 583-594出版社
WILEY
DOI: 10.1111/mpp.13178
关键词
bacterial pathogen; genome editing; rice; type I-C CRISPR-Cas system; Xanthomonas oryzae pv; oryzae
资金
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDA24020101, XDPB16]
- National Natural Science Foundation of China [32001891, 31901868]
Efficient genome editing using an identified type I-C CRISPR-Cas system in Xoo allows for precise modifications of the genome, providing a valuable tool for functional genomics studies of rice bacterial blight and potentially other CRISPR-harboring bacterial plant pathogens.
Efficient and modular genome editing technologies that manipulate the genome of bacterial pathogens will facilitate the study of pathogenesis mechanisms. However, such methods are yet to be established for Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of rice bacterial blight. We identified a single type I-C CRISPR-Cas system in the Xoo genome and leveraged this endogenous defence system for high-efficiency genome editing in Xoo. Specifically, we developed plasmid components carrying a mini-CRISPR array, donor DNA, and a phage-derived recombination system to enable the efficient and programmable genome editing of precise deletions, insertions, base substitutions, and gene replacements. Furthermore, the type I-C CRISPR-Cas system of Xoo cleaves target DNA unidirectionally, and this can be harnessed to generate large genomic deletions up to 212 kb efficiently. Therefore, the genome-editing strategy we have developed can serve as an excellent tool for functional genomics of Xoo, and should also be applicable to other CRISPR-harbouring bacterial plant pathogens.
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