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

• Published in: Communications biology Vol. 7; no. 1; pp. 1129 - 11
• Main Authors: Li, Feng-Yu, Tan, Xin-Ee, Shimamori, Yuzuki, Kiga, Kotaro, Veeranarayanan, Srivani, Watanabe, Shinya, Nishikawa, Yutaro, Aiba, Yoshifumi, Sato’o, Yusuke, Miyanaga, Kazuhiko, Sasahara, Teppei, Hossain, Sarah, Thitiananpakorn, Kanate, Kawaguchi, Tomofumi, Nguyen, Huong Minh, Yeo Syin Lian, Adeline, Sultana, Sharmin, Alessa, Ola, Kumwenda, Geoffrey, Sarangi, Jayathilake, Revilleza, Jastin Edrian Cocuangco, Baranwal, Priyanka, Faruk, Mohammad Omar, Hidaka, Yuya, Thu, Myat, Arbaah, Mahmoud, Batbold, Anujin, Maniruzzaman, Liu, Yi, Duyen, Ho Thi My, Sugano, Takashi, Tergel, Nayanjin, Shimojyo, Takayuki, Cui, Longzhu
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 13.09.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 42; 42/41; 45; 45/41; 631/326/1320; 631/326/22/1434; Anti-Bacterial Agents - pharmacology; Antibiotic resistance; Antibiotics; Antimicrobial agents; Bactericidal activity; Biomedical and Life Sciences; Capsid - drug effects; Capsid - metabolism; Capsids; Contamination; Copy number; CRISPR; CRISPR-Cas Systems; Drug resistance; Gene deletion; Life Sciences; Methicillin; Methicillin-Resistant Staphylococcus aureus - drug effects; Methicillin-Resistant Staphylococcus aureus - genetics; Multidrug resistance; Pathogens; Phages; Staphylococcal Infections - drug therapy; Staphylococcal Infections - microbiology; Staphylococcus aureus; Staphylococcus infections
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-024-06754-w

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. Phagemid-based capsid system delivers CRISPR-Cas13a to target MRSA, showing efficient packaging, minimal wild-type phage contamination, and specific bactericidal activity, providing a promising alternative to antibiotics.