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- W3091859545 abstract "ABSTRACT Purple non-sulfur photosynthetic bacteria (PNSB) such as R. 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 both genome editing and gene down-regulation in R. capsulatus . About 90% editing efficiency was achieved by using CRISPR/Cas12a driven by a strong promoter P puc when targeting ccoO or nifH gene. When both genes were simultaneously targeted, the multiplex gene editing efficiency reached >63%. In addition, CRISPR interference using deactivated Cas12a was also evaluated using reporter genes gfp and lacZ , and the repression efficiency reached >80%. In summary, our work represents the first report to develop CRISPR/Cas12a mediated genome editing/transcriptional repression in R. capsulatus , which would greatly accelerate PNSB-related researches. IMPORTANCE Purple non-sulfur photosynthetic bacteria (PNSB) such as R. capsulatus serve as a versatile platform for fundamental studies and various biotechnological applications. However, lack of efficient gene editing tools remains a main obstacle for progressing in PNSB-related researches. Here, we developed CRISPR/Cas12a for genome editing via the non-homologous end joining (NHEJ) repair machinery in R. capsulatus . In addition, DNase-deactivated Cas12a was found to simultaneously suppress multiple targeted genes. Taken together, our work offers a new set of tools for efficient genome engineering in PNSB such as R. capsulatus ." @default.
- W3091859545 created "2020-10-15" @default.
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- W3091859545 date "2020-10-06" @default.
- W3091859545 modified "2023-09-25" @default.
- W3091859545 title "CRISPR/Cas12a mediated genome engineering in photosynthetic bacteria" @default.
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- W3091859545 doi "https://doi.org/10.1101/2020.10.05.327569" @default.
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