Mutation-induced penicillin binding protein 2A inactivation through CRISPR-dCas9 gene editing in methicillin-resistant S. aureus
4 affiliations
- 1Institute of Molecular Biology and Biotechnology, University of Lahore, Lahore, Pakistan
- 2Institute-National Research Institute, Radzikow, Blonie, Poland
- 3Department of Pediatrics, Punjab Institute of Cardiology, Lahore, Pakistan
- 4Centre for Research in Molecular Medicine (CRiMM), University of Lahore, Lahore, Pakistan
Abstract
Background and Objectives:: The global rise of antimicrobial resistance threatens effective treatment of bacterial infections. Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen whose resistance to β-lactam antibiotics is primarily mediated by the mecA gene, encoding the low-affinity penicillin-binding protein PBP2a. This study aimed to investigate whether CRISPR-dCas9-mediated targeting of mecA could suppress resistance-associated gene expression and restore β-lactam susceptibility in MRSA.
Materials and Methods:: Seven target sites within mecA, comprising two non-coding and five coding regions, were selected for CRISPR-dCas9 targeting. The effects of gene interference were evaluated using antibiotic disk diffusion assays, minimum inhibitory concentration (MIC) testing with gradient E-strips, RT-PCR, and Sanger sequencing.
Results:: Targeting non-coding regions suppressed mecA transcription and reduced PBP2a production, whereas targeting coding regions generated mutations associated with dysfunctional PBP2a. CRISPR-dCas9 treatment significantly reduced mecA expression and increased MRSA susceptibility to β-lactam antibiotics, particularly ampicillin and cefixime. MIC values decreased by up to 12-fold and 6-fold, respectively (p<0.05).
Conclusion:: CRISPR-dCas9-mediated targeting of mecA effectively reduced β-lactam resistance in MRSA. This approach demonstrates potential for precision-based antimicrobial resistance management and warrants further investigation as a strategy for combating drug-resistant bacterial infections.
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BibTeX
@article{khan2026mutationinduced,
title = {{Mutation-induced penicillin binding protein 2A inactivation through CRISPR-dCas9 gene editing in methicillin-resistant S. aureus}},
author = {Arooj Khan and Adnan Iqbal and Husnain Athar and Ahsan Sattar Sheikh},
journal = {Iranian Journal of Microbiology},
year = {2026},
month = nov,
doi = {10.18502/ijm.v18i5.22865},
url = {https://doi.org/10.18502/ijm.v18i5.22865},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Mutation-induced penicillin binding protein 2A inactivation through CRISPR-dCas9 gene editing in methicillin-resistant S. aureus
AU - Arooj Khan
AU - Adnan Iqbal
AU - Husnain Athar
AU - Ahsan Sattar Sheikh
PY - 2026
DA - 2026/11/06
JO - Iranian Journal of Microbiology
DO - 10.18502/ijm.v18i5.22865
UR - https://doi.org/10.18502/ijm.v18i5.22865
AB - Background and Objectives:: The global rise of antimicrobial resistance threatens effective treatment of bacterial infections. Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen whose resistance to β-lactam antibiotics is primarily mediated by the mecA gene, encoding the low-affinity penicillin-binding protein PBP2a. This study aimed to investigate whether CRISPR-dCas9-mediated targeting of mecA could suppress resistance-associated gene expression and restore β-lactam susceptibility in MRSA. Materials and Methods:: Seven target sites within mecA, comprising two non-coding and five coding regions, were selected for CRISPR-dCas9 targeting. The effects of gene interference were evaluated using antibiotic disk diffusion assays, minimum inhibitory concentration (MIC) testing with gradient E-strips, RT-PCR, and Sanger sequencing. Results:: Targeting non-coding regions suppressed mecA transcription and reduced PBP2a production, whereas targeting coding regions generated mutations associated with dysfunctional PBP2a. CRISPR-dCas9 treatment significantly reduced mecA expression and increased MRSA susceptibility to β-lactam antibiotics, particularly ampicillin and cefixime. MIC values decreased by up to 12-fold and 6-fold, respectively (p<0.05). Conclusion:: CRISPR-dCas9-mediated targeting of mecA effectively reduced β-lactam resistance in MRSA. This approach demonstrates potential for precision-based antimicrobial resistance management and warrants further investigation as a strategy for combating drug-resistant bacterial infections.
SN - 2008-3289
SN - 2008-4447
ER -