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Article Abstract

In response to the escalating antibiotic resistance in multidrug-resistant pathogens, we propose an innovative phagemid-based capsid system to generate CRISPR-Cas13a-loaded antibacterial capsids (AB-capsids) for targeted therapy against multidrug-resistant Staphylococcus aureus. Our optimized phagemid system maximizes AB-capsid yield and purity, showing a positive correlation with phagemid copy number. Notably, an 8.65-fold increase in copy number results in a 2.54-fold rise in AB-capsid generation. Phagemids carrying terL-terS-rinA-rinB (prophage-encoded packaging site genes) consistently exhibit high packaging efficiency, and the generation of AB-capsids using lysogenized hosts with terL-terS deletion resulted in comparatively lower level of wild-type phage contamination, with minimal compromise on AB-capsid yield. These generated AB-capsids selectively eliminate S. aureus strains carrying the target gene while sparing non-target strains. In conclusion, our phagemid-based capsid system stands as a promising avenue for developing sequence-specific bactericidal agents, offering a streamlined approach to combat antibiotic-resistant pathogens within the constraints of efficient production and targeted efficacy.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11399120PMC
http://dx.doi.org/10.1038/s42003-024-06754-wDOI Listing

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Phagemid-based capsid system for CRISPR-Cas13a antimicrobials targeting methicillin-resistant Staphylococcus aureus.

Commun Biol

September 2024

Division of Bacteriology, Department of Infection and Immunity, School of Medicine, Jichi Medical University, Shimotsuke city, Tochigi, 329-0498, Japan.

Article Synopsis
  • The study presents a new phagemid-based system that produces CRISPR-Cas13a-loaded antibacterial capsids (AB-capsids) specifically targeting multidrug-resistant Staphylococcus aureus.
  • By optimizing phagemid copy numbers, researchers achieved higher yields and purity of AB-capsids, illustrating a direct relationship between phagemid quantity and capsid production.
  • The developed AB-capsids effectively eliminate targeted S. aureus strains while leaving non-target strains unharmed, showcasing their potential as effective tools against antibiotic-resistant bacteria.
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