Innovative Delivery System Combining CRISPR-Cas12f for Combatting Antimicrobial Resistance in Gram-Negative Bacteria

• DOI: 10.1021/acssynbio.4c00112
• Publication Date: 2024-06-12T06:59:45Z
• Authors: Teng-Fei Long; Shi-Ying Zhou; Zi-Lei Huang; Gong Li; Qin Zhong; Xiao-Jing Zhang; Yuan-Yuan Li; Cai-Ping Chen; Li-Juan Xia; Ran Wei; Lei Wan; Ang Gao; Hao Ren; Xiao-Ping Liao; Ya-Hong Liu; Liang Chen; Jian Sun
• Keywords: Bacterial conjugation

Antimicrobial resistance poses a significant global challenge, demanding innovative approaches, such as the CRISPR-Cas-mediated plasmid or gene-curing system, to effectively combat this urgent crisis. To enable successful curing of antimicrobial genes plasmids through CRISPR-Cas technology, development an efficient broad-host-range delivery system is paramount. In study, we have successfully designed and constructed novel functional gene plasmid, pQ-mini, utilizing backbone Inc.Q plasmid. Moreover, integrated CRISPR-Cas12f into pQ-mini in broad-host bacteria. Our findings demonstrate that facilitates highly transfer genetic elements diverse bacteria, particularly various species order Enterobacterales, exhibiting broader host range superior conjugation efficiency compared commonly used pMB1-like Notably, delivers antimicrobial-resistant strains, resulting remarkable efficiencies for plasmid-borne mcr-1 blaKPC are comparable those achieved by previously reported pCasCure system. conclusion, our study establishes optimizes vector. Furthermore, combination with demonstrates its potential delivery, highlighting promising role tool against growing threat resistance.