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Updated: Jun 23, 2026

Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira
14:49

Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira

Published on: August 14, 2021

Lipoplex as a useful tool for resensitization of methicillin-resistant Staphylococcus aureus to erythromycin via

Aysegul Ates1, Melis Onal2, Mert Senyigit2

  • 1Pharmaceutical Microbiology Department, Faculty of Pharmacy, Ege University, 35040 Izmir, Turkiye.

International Journal of Pharmaceutics
|June 20, 2026
PubMed

Insights

CRISPR-Cas9 gene editing delivered via lipoplexes successfully reduced erythromycin resistance in MRSA. This approach resensitized bacteria to antibiotics, offering a promising strategy against antimicrobial resistance.

Area of Science:

  • Molecular Biology
  • Gene Editing Technologies
  • Antimicrobial Resistance

Background:

  • Antimicrobial resistance (AMR) is a growing global health threat, with Methicillin-resistant Staphylococcus aureus (MRSA) posing significant clinical challenges.
  • Novel therapeutic strategies are crucial to combat resistant bacterial infections.

Purpose of the Study:

  • To investigate the efficacy of CRISPR-Cas9 gene editing for targeting the ermA gene responsible for erythromycin resistance in MRSA.
  • To develop and characterize a liposome-based delivery system for CRISPR-Cas9 components.

Main Methods:

  • Development of cationic liposomes (Lips) for pCasSA plasmid delivery.
  • Formation of lipoplexes (Lpx) through electrostatic interaction between Lips and pCasSA.
  • Assessment of Lpx/pCasSA cytotoxicity in L929 cells.
  • Confirmation of gene editing and ermA expression reduction using Sanger sequencing and RT-qPCR, enhanced by sonoporation.
  • Evaluation of restored antibiotic susceptibility through phenotypic assays (MIC and inhibition zones).

Main Results:

  • Stable liposomes (<200 nm, PDI < 0.3, ZP > +30 mV) were successfully formulated.
  • Lipoplex formulations exhibited low cytotoxicity.
  • CRISPR-Cas9 delivery via Lpx combined with sonoporation achieved a 75% reduction in ermA gene expression.
  • Significant restoration of erythromycin susceptibility was observed, with an 8-fold decrease in MIC and a 2.5-fold increase in inhibition zones.

Conclusions:

  • Liposome-mediated CRISPR-Cas9 delivery is an effective strategy for combating AMR.
  • This approach shows potential for resensitizing MRSA to existing antibiotics, thereby restoring their clinical efficacy.
Keywords:
Antimicrobial resistanceCRISPR-Cas9Gene editingLipoplexMethicillin-resistant Staphylococcus aureusResensitization

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