Efficient synthesis of CRISPR-Cas13a-antimicrobial capsids against MRSA facilitated by silent mutation incorporation.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
13 Jul 2024
13 Jul 2024
Historique:
received:
26
12
2023
accepted:
09
07
2024
medline:
14
7
2024
pubmed:
14
7
2024
entrez:
13
7
2024
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 39003336
doi: 10.1038/s41598-024-67193-5
pii: 10.1038/s41598-024-67193-5
doi:
Substances chimiques
Anti-Bacterial Agents
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
16225Subventions
Organisme : JSPS KAKENHI
ID : 22K15673
Organisme : JSPS KAKENHI
ID : 22K19386
Organisme : JSPS KAKENHI
ID : 21K19666
Organisme : JSPS KAKENHI
ID : 21K19488
Organisme : Japan Agency for Medical Research and Development
ID : JP21fk0108497
Organisme : Japan Agency for Medical Research and Development
ID : JP23wm0325065
Organisme : Japan Agency for Medical Research and Development
ID : JP21ae0121045
Organisme : Moonshot Research and Development Program
ID : JPJ009237
Informations de copyright
© 2024. The Author(s).
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