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A Transposon-Associated CRISPR/Cas9 System Specifically Eliminates both Chromosomal and Plasmid-Borne mcr-1 in

Yu-Zhang He1,2,3,4, Jin-Ru Yan1,2,3, Bing He1,2,3

  • 1National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.

Antimicrobial Agents and Chemotherapy
|August 2, 2021
PubMed

Insights

A novel CRISPR/Cas9 system effectively eliminates the mcr-1 gene, a colistin resistance factor, from bacteria. This approach restores colistin sensitivity and prevents further spread of antimicrobial resistance.

Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Antimicrobial resistance, particularly to last-resort antibiotics like colistin, is a major global public health threat.
  • The mcr-1 gene, conferring colistin resistance, is rapidly spreading among Enterobacteriaceae species, often facilitated by mobile genetic elements like transposons.
  • Existing treatments are becoming less effective against resistant bacterial strains.

Purpose of the Study:

  • To develop a targeted gene-editing strategy to eliminate the mcr-1 gene from bacteria.
  • To assess the efficacy of a CRISPR/Cas9 system in combating colistin resistance.
  • To investigate the potential of this system as a therapeutic approach against antimicrobial resistance.

Main Methods:

  • Development of a suicide plasmid carrying an ISApl1-flanked CRISPR/Cas9 system.
  • Application of the system to target and eliminate both plasmid-borne and chromosome-borne mcr-1 genes.
  • Evaluation of bacterial sensitivity to colistin post-treatment.
  • Assessment of immunity against exogenous mcr-1 acquisition.

Main Results:

  • The CRISPR/Cas9 system successfully removed the mcr-1 gene from Escherichia coli, restoring colistin sensitivity.
  • The system demonstrated efficacy against both plasmid-mediated and chromosomally-located mcr-1.
  • Treated bacteria exhibited resistance to acquiring new mcr-1 genes via plasmids.
  • High efficiency in gene curing and bacterial eradication was observed.

Conclusions:

  • The transposon-associated CRISPR/Cas9 system is a potent tool for eradicating the mcr-1 gene.
  • This technology offers a promising therapeutic strategy to combat colistin resistance and control the spread of antimicrobial resistance in clinical settings.
Keywords:
CRISPR/Cas9ISApl1antibiotic resistancemcr-1transposon

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