Optimizing genome editing efficiency in <i>Streptomyces fradiae</i> via a CRISPR/Cas9n-mediated editing system.

ABSTRACT: Streptomyces fradiae is an important bioresource to produce various antibacterial natural products, however, the time-consuming and labor-intensive genome editing toolkits hindered the construction and application of engineered strains, and this study aimed to establish an efficient CRISPR/Cas9n genome editing system in S. fradiae. Initially, the CRISPR/Cas9-mediated editing tool was employed to replace those awkward genome editing tools that relied on homologous recombination, while the off-target Cas9 exhibited high toxicity to S. fradiae Sf01. Therefore, the nickase mutation D10A, high-fidelity mutations including N497A, R661A, Q695A, and Q926A, and thiostrepton-induced promotor P_tipA were incorporated into the Cas9 expression cassette, which reduc

SUBMITTER: Wu Y 

PROVIDER: S-EPMC11837490 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature