Implementation of a mycobacterial CRISPRi platform in Mycobacterium abscessus and demonstration of the essentiality of ftsZ_Mab.

ABSTRACT: Mycobacterium abscessus (Mab) is a highly drug-resistant non-tuberculous mycobacterial species that causes debilitating TB-like pulmonary infections. The lack of genetic tools has hampered characterization of its extensive repertoire of virulence factors, antimicrobial resistance mechanisms, and drug targets. In this study, we evaluated the performance of a mycobacterial single plasmid CRISPRi-dCas9 system optimized for M. tuberculosis and M. smegmatis for inducible gene silencing in Mab. The efficacy of CRISPRi-mediated repression of two antibiotic resistance genes (bla_Mab, whiB7_Mab) and two putative essential genes (ftsZ_Mab,topA_Mab) was determined by measuring mRNA transcript levels and phenotypic outcomes. While our results support the utility

SUBMITTER: Gupta R 

PROVIDER: S-EPMC11552333 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature