期刊
MICROBIAL BIOTECHNOLOGY
卷 14, 期 6, 页码 2700-2710出版社
WILEY
DOI: 10.1111/1751-7915.13805
关键词
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资金
- Xiamen University [0660-X2123310]
- Natural Science Foundation of Fujian Province of China [2020J05011]
- XMU Training Program of Innovation and Entrepreneurship for Undergraduates [2020Y1000, 202010384194]
- ZhenSheng Biotech, China
The study developed a CRISPR/Cas12a system for genome editing and transcriptional regulation in R. capsulatus, achieving high editing efficiency and offering broad potential applications.
Purple non-sulfur photosynthetic bacteria (PNSB) such as Rhodobacter capsulatus serve as a versatile platform for fundamental studies and various biotechnological applications. In this study, we sought to develop the class II RNA-guided CRISPR/Cas12a system from Francisella novicida for genome editing and transcriptional regulation in R. capsulatus. Template-free disruption method mediated by CRISPR/Cas12a reached - 90% editing efficiency when targeting ccoO or nifH gene. When both genes were simultaneously edited, the multiplex editing efficiency reached > 63%. In addition, CRISPR interference (CRISPRi) using deactivated Cas12a was also evaluated using reporter genes egfp and IacZ, and the transcriptional repression efficiency reached similar to 80%. In summary, our work represents the first report to develop CRISPR/Cas12a-mediated genome editing and transcriptional regulation in R. capsulatus, which would greatly accelerate PNSB-related researches.
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