A CRISPR/Cas9-based system using dual-sgRNAs for efficient gene deletion in <i>Mycobacterium abscessus</i>.

ABSTRACT: The increasing global prevalence of Mycobacterium abscessus infections presents a significant clinical challenge due to the pathogen's intrinsic resistance to multiple antibiotics and poor treatment outcomes. Despite the necessity of genetic tools for studying its physiology, pathogenesis, and drug resistance, efficient methods for large-fragment deletions remain underdeveloped. Here, we report a CRISPR/Cas9-based dual-sgRNA system employing Streptococcus thermophilus CRISPR1-Cas9 (Sth1Cas9), enabling efficient large-fragment knockout in M. abscessus with deletion efficiencies exceeding 90% at certain loci and spanning up to 16.7 kb. Furthermore, we systematically optimized the modular arrangement of genetic components in Cas9/dual-sgRNA expression plasmids and refined

SUBMITTER: Li L 

PROVIDER: S-EPMC12283748 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature