铜绿假单胞菌是烧、创伤感染的重要病原菌，多重耐药铜绿假单胞菌的广泛流行已严重威胁了烧伤患者的生命安全。对铜绿假单胞菌必需基因进行深入研究，不仅有助于了解细菌的生长、毒力等基本特性，也有助于筛选新的抗菌药物靶标。CRISPRi技术是一种高效地研究细菌必需基因的新策略，具有操作简便、通量高、效率高以及可诱导控制等优点。前期，已在铜绿假单胞菌中建立了CRISPRi系统，并用该系统研究两个必需基因。本研究计划首先运用已建立的CRISPRi系统构建一个靶向铜绿假单胞菌PAO1的所有必需基因的CRISPRi文库，建立一个高效PAO1必需基因研究平台。其次，通过高通量的方法，对PAO1的352个必需基因进行表型研究，重点对46个未知功能基因进行深入研究。最后，基于上述结果寻找一些潜在的抗生素作用靶点。本研究的实施可为铜绿假单胞菌必需基因研究和抗生素等靶点筛选提供一个有力工具。

Pesudomonas aeruginosa is one of the important pathogens in burns and trauma ward. With the widely distribution of multi-drug resistance, P. aeruginosa become a big threat to burn patients. Investigation of P. aeruginosa essential genes promote further understanding of bacteria growth, cytotoxicity and so on. It can also help to identify new potential antibiotic targets. With the advantages of easy-to-operation, high thoughput and high efficiency, CRISPRi technology is becoming a new efficient method to study essential genes in bacterium. In previous work, we built a CRISPRi system in P. aeruginosa and the expression of two essential genes ( PA0611 and PA0715) were repressed with this CRIPSRi system. In this study, we first tried to build a CRISPRi library which can target all 352 essential genes of PAO1. With the CRISPRi library, we further investigate the gene function of each essential gene by testing phenotype changes and gene expression changes, especially the 46 function-unknown essential genes. Last, search potential antibiotic targets based on the above results. With the project, we will build a CRISPRi library targeting all essential genes. The success of this study could serve as a powerful tool for essential gene investigation and antibiotic exploration in P. aeruginosa.