Scholarlink

<p>Metallo-β-lactamases (MBLs), such as those encoded by <i>bla</i><sub>IMP-1</sub>, confer resistance to carbapenem antibiotics and represent a critical challenge in treating infections caused by multidrug-resistant <i>Pseudomonas aeruginosa</i>. Here, we report a programmable antimicrobial strategy that restores bacterial antibiotic susceptibility through phage capsid-mediated delivery of CRISPR-Cas13a. We engineered a non-replicative phage capsid, which we called antibacterial capsid (AB-Capsid), packaged with a phagemid encoding a codon-optimized Cas13a from <i>Leptotrichia shahii</i> (<i>cas13aPA</i>) and a guide RNA targeting <i>bla</i><sub>IMP-1</sub>. The resulting construct, AB-Capsid_<i>cas13aPA</i>_<i>bla</i><sub>IMP-1</sub>, specifically inhibited the growth of <i>bla</i><sub>IMP-1</sub>-expressing <i>P. aeruginosa</i> and significantly reduced the minimum inhibitory concentration (MIC) of imipenem. No bactericidal effect was observed in the absence of the target gene or with a non-targeting AB-Capsid. Furthermore, spacer-dependent and expression-level-dependent killing activity was confirmed using inducible <i>bla</i><sub>IMP-1</sub> systems. These findings demonstrate that programmable AB-Capsids delivering Cas13a provide a gene-specific, non-replicative antimicrobial platform capable of reversing drug resistance and represent a versatile class of CRISPR-based antibiotic adjuvants.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Gene-specific reversal of carbapenem-resistant Pseudomonas aeruginosa via phage-delivered CRISPR-Cas13a

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年12月16日

查看全文
  • Tomofumi Kawaguchi,
  • Shinya Watanabe,
  • Yi Liu,
  • Yoshifumi Aiba,
  • Xin-Ee Tan,
  • Srivani Veeranarayanan,
  • Kazuhiko Miyanaga,
  • Teppei Sasahara,
  • Yuzuki Shimamori,
  • Ola Alessa,
  • Yuya Hidaka,
  • Myat Thu,
  • Orawee Kaewprasert,
  • Varsha Rani,
  • Md Razib Hossain,
  • Vivekanandan Palaninathan,
  • Palanichamy Esakkiraj,
  • Taufik Fatwa Nur Hakim,
  • Karthik Maruthan,
  • Pedro B. Fernandes,
  • Mahmoud Arbaah,
  • Anujin Batbold,
  • Maniruzzaman,
  • Sarah Hossain,
  • Takashi Sugano,
  • Hidetaka Uematsu,
  • Dhammika Leshan Wannigama,
  • Kotaro Kiga,
  • Longzhu Cui

摘要

Metallo-β-lactamases (MBLs), such as those encoded by blaIMP-1, confer resistance to carbapenem antibiotics and represent a critical challenge in treating infections caused by multidrug-resistant Pseudomonas aeruginosa. Here, we report a programmable antimicrobial strategy that restores bacterial antibiotic susceptibility through phage capsid-mediated delivery of CRISPR-Cas13a. We engineered a non-replicative phage capsid, which we called antibacterial capsid (AB-Capsid), packaged with a phagemid encoding a codon-optimized Cas13a from Leptotrichia shahii (cas13aPA) and a guide RNA targeting blaIMP-1. The resulting construct, AB-Capsid_cas13aPA_blaIMP-1, specifically inhibited the growth of blaIMP-1-expressing P. aeruginosa and significantly reduced the minimum inhibitory concentration (MIC) of imipenem. No bactericidal effect was observed in the absence of the target gene or with a non-targeting AB-Capsid. Furthermore, spacer-dependent and expression-level-dependent killing activity was confirmed using inducible blaIMP-1 systems. These findings demonstrate that programmable AB-Capsids delivering Cas13a provide a gene-specific, non-replicative antimicrobial platform capable of reversing drug resistance and represent a versatile class of CRISPR-based antibiotic adjuvants.