AI Article Synopsis

  • The lab created a phagemid system to generate CRISPR-Cas13a-antimicrobial capsids specifically targeting MRSA to combat rising antimicrobial resistance.
  • They faced a challenge with unwanted wild-type phage production during packaging, which was addressed by introducing silent mutations to reduce contamination while maintaining efficiency.
  • The optimized system showed effective sequence-specific killing of MRSA strains but highlights the need for further research on its effectiveness against other bacteria and in real body conditions.

Article Abstract

In response to the escalating global threat of antimicrobial resistance, our laboratory has established a phagemid packaging system for the generation of CRISPR-Cas13a-antimicrobial capsids targeting methicillin-resistant Staphylococcus aureus (MRSA). However, a significant challenge arose during the packaging process: the unintentional production of wild-type phages alongside the antimicrobial capsids. To address this issue, the phagemid packaging system was optimized by strategically incorporated silent mutations. This approach effectively minimized contamination risks without compromising packaging efficiency. The study identified the indispensable role of phage packaging genes, particularly terL-terS, in efficient phagemid packaging. Additionally, the elimination of homologous sequences between the phagemid and wild-type phage genome was crucial in preventing wild-type phage contamination. The optimized phagemid-LSAB(mosaic) demonstrated sequence-specific killing, efficiently eliminating MRSA strains carrying target antibiotic-resistant genes. While acknowledging the need for further exploration across bacterial species and in vivo validation, this refined phagemid packaging system offers a valuable advancement in the development of CRISPR-Cas13a-based antimicrobials, shedding light on potential solutions in the ongoing battle against bacterial infections.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11246472PMC
http://dx.doi.org/10.1038/s41598-024-67193-5DOI Listing

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