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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

References

    1. Cowman S, van Ingen J, Griffith DE, Loebinger MR. 2019. Non-tuberculous mycobacterial pulmonary disease. Eur Respir J 54:1900250. doi:10.1183/13993003.00250-2019. - DOI - PubMed
    1. Johansen MD, Herrmann JL, Kremer L. 2020. Non-tuberculous mycobacteria and the rise of Mycobacterium abscessus. Nat Rev Microbiol 18:392–407. doi:10.1038/s41579-020-0331-1. - DOI - PubMed
    1. Lee MR, Sheng WH, Hung CC, Yu CJ, Lee LN, Hsueh PR. 2015. Mycobacterium abscessus complex infections in humans. Emerg Infect Dis 21:1638–1646. doi:10.3201/2109.141634. - DOI - PMC - PubMed
    1. Koh WJ, Jeong BH, Kim SY, Jeon K, Park KU, Jhun BW, Lee H, Park HY, Kim DH, Huh HJ, Ki CS, Lee NY, Kim HK, Choi YS, Kim J, Lee SH, Kim CK, Shin SJ, Daley CL, Kim H, Kwon OJ. 2017. Mycobacterial characteristics and treatment outcomes in Mycobacterium abscessus lung disease. Clin Infect Dis 64:309–316. doi:10.1093/cid/ciw724. - DOI - PubMed
    1. Koh WJ, Stout JE, Yew WW. 2014. Advances in the management of pulmonary disease due to Mycobacterium abscessus complex. Int J Tuber Lung Dis 18:1141–1148. doi:10.5588/ijtld.14.0134. - DOI - PubMed

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