FEMS microbiology letters

Using CRISPR-Cas to Design DNA Tools Against Drug-Resistant Klebsiella Pneumoniae Bacteria

Updated

Abstract

All 5 clinical isolates of multidrug-resistant Klebsiella pneumoniae demonstrated resistance to multiple antibiotics, with minimum inhibitory concentrations (MICs) ranging from 4-128 μg/mL for imipenem.

  • Resistance gene analysis showed the presence of blaOXA-48-like and blaCTX-M-15 in all isolates, with blaNDM-1 found in one isolate.
  • Two CRISPR-based plasmid systems were designed for potential genome editing applications.
  • Apramycin at 50 μg/mL was identified as a suitable selection marker for plasmid construction.
  • The pSGKP-AmpR(Pro)-ApmR plasmid was successfully constructed, while cloning of Cas9 and APOBEC constructs faced technical challenges.
  • The findings suggest a need for isolate-specific plasmid design and optimization of guide RNA sequences.

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Full Text

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Funding

Competing interests

None declared.
PubMed

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