利用噬菌体和纳米材料载体递呈CRISPR/Cas9系统靶向清除细菌耐药性研究

细菌耐药严重影响了抗生素对细菌感染治疗的效果。近年来，噬菌体抗菌技术备受关注，同时发展了噬菌体递呈CRISPR/Cas9系统靶向细菌基因组以杀死细菌，但上述技术还存在诸多问题，例如容易产生抗性且杀菌效力尚显不足、无法抑制耐药质粒转移等。本研究将新建一步法噬菌体重组CRISPR/Cas9系统技术，递呈CRISPR/Cas9系统靶向清除细菌耐药质粒以恢复抗生素敏感性，并联合使用致敏抗生素有效治疗耐药菌感染并阻断耐药质粒转移；同时建立纳米载体递呈CRISPR/Cas9系统技术，实现跨菌种清除细菌耐药性；还将系统阐明CRISPR/Cas9不同递呈系统效率、耐药性清除机制、联合抗生素治疗耐药菌感染效果等关键问题。本研究提出CRISPR/Cas9系统清除耐药性并联合致敏抗生素治疗耐药菌感染的策略，并通过体内外实验验证该方法的疗效，为防治细菌耐药提供理论和技术上的支撑。

Bacterial antibiotic-resistance seriously defected efficacy of antibiotics. Recently, researchers highly focused on phage therapy and developed CRISPR/Cas-integrated phage to kill bacteria through lethal genome targeting. However, many problems remained in previous work, such as development of new resistance, insufficient bactericidal efficacy and being incapable of preventing the transfer of resistance plasmid. Our study invent an one-step method for phage genome recombination with the CRISPR/Cas9 and delivery the CRISPR/Cas9 system to target bacterial resistance plasmids, thus to restore sensitivity of common antibiotics and block the horizontal transfer of bacterial antibiotic resistance genes. We will also develop nano-materials-based vectors to deliver the CRISPR/Cas9 system within broader range of bacteria strain and species. Delivery efficiency of the CRISPR/Cas9, clearance rate of resistance plasmid and combined treatment effect with resensitized antibiotic will be comprehensively illustrated. Through proposing the strategy of removing bacterial resistance using CRISPR/Cas9 and verifying combined therapeutic efficacy against resistant bacteria in vitro and in vivo, this research will lay the theoretical and technical foundation for treatment and prevention of drug-resistant pathogens.