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. 2021 Aug 14;49(16):9594–9605. doi: 10.1093/nar/gkab673

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Construction and characterization of CRISPR-CDA-Cas9-mediated base editor system in B. subtilis. (A) Design and construction of CRISPR-CDA-Cas9 mediating base editors. The fusion protein comprising nCas9/dCas9 and CDA under the control of xylose promoter P_xylA was integrated into the lacA locus in B. subtilis. The repressor gene xylR was constitutively expressed in the expression cassette. A strong constitutive promoter P43 controlled the designed sgRNA. (B) Workflow for gene editing identification and verification at the single-clone level and population level in B. subtilis. (C) The editing efficiency of CRISPR-CDA-dCas9 and CRISPR-CDA-nCas9 at sigE site. (D) Base conversion targeting sigE at the single-clone level. Thirty colonies were selected and subjected to gene sequencing and the conversion efficiency was calculated. The PAM sequence is blue, whereas the modified bases are red.