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. 2022 Apr 19;66(4):e0009322.
doi: 10.1128/aac.00093-22. Epub 2022 Mar 21.

CRISPR Inhibition of Essential Peptidoglycan Biosynthesis Genes in Mycobacterium abscessus and Its Impact on β-Lactam Susceptibility

Natalia Kurepina  1 , Liang Chen  1   2 , Kaelea Composto  1 , Dalin Rifat  3 , Eric L Nuermberger  3 , Barry N Kreiswirth  1   2
Affiliations

CRISPR Inhibition of Essential Peptidoglycan Biosynthesis Genes in Mycobacterium abscessus and Its Impact on β-Lactam Susceptibility

Natalia Kurepina et al. Antimicrob Agents Chemother. .

Abstract

We utilized a CRISPR interference (CRISPRi) assay to control the gene expressions of two predicted essential peptidoglycan biosynthesis genes, pbpB and cwIM, in Mycobacterium abscessus and to evaluate their contribution to β-lactam susceptibility. Our results showed that CRISPR inhibition of each gene led to a significant 3-log10 reduction in CFU in the presence of imipenem but not for cefoxitin. These results demonstrate that CRISPRi provides an experimental approach to study drug/target interactions in M. abscessus.

Keywords: CRISPRi; Mycobacterium abscessus; peptidoglycan biosynthesis; β-lactam.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIG 1
FIG 1
The growth of M. abscessus ATCC 19977 CRISPRi constructs exposed to different ATc concentrations.
FIG 2
FIG 2
The interaction of ATc and either imipenem or cefoxitin and their effects on the growth of M. abscessus ATCC 19977 CRISPRi constructs.
See this image and copyright information in PMC

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