A Strictly Inducible and Orthogonal Dre-rox System for Precise and Markerless Genome Editing in <i>Bacillus subtilis</i>.

ABSTRACT: Site-specific recombination enables precise and modular genome engineering in microbial systems. In Bacillus subtilis, Cre is the most commonly used site-specific recombinase (SSR) and has been widely applied in genome engineering. Developing SSRs with comparable performance to Cre that can also function orthogonally would significantly expand the genome engineering toolkit. We established a resistance gene-based reporter in B. subtilis to assess the genome editing potential of the Dre-rox system. A theophylline-inducible riboswitch tightly controlled Dre expression to minimize leaky recombination, improving the specificity of rox-mediated recombination. Notably, Dre and Cre function without crosstalk at their respective recognition sites. This orthogonal combination enabled

SUBMITTER: Lv J 

PROVIDER: S-EPMC12331466 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature