A CRISPRi screen of essential genes of Neisseria gonorrhoeae

摘要 淋病奈瑟菌是性传播感染淋病的唯一病原体， 只存在于人类中N.淋病一直与人类宿主共同进化 现代历史，并已演变出与其发病模式相关的特定发病机制。 传播和它殖民的解剖部位。淋病的发病率沿着快速的 获得抗生素耐药性。因此迫切需要了解N. 淋病发病机制，以开发新的治疗方案或候选疫苗。N.淋病 是天然的DNA转化能力，是少数几种细菌之一， 规范权限。这项资助将使用CRISPR干扰技术（CRISPRi）来确定 N.淋病，并提供研究社区的能力， 必要的基因。这项工作将补充我们资助的R24资源赠款， 每个开放阅读框的插入突变体。淋病，但不能突变的基本 基因. R24项目正在利用N.淋病和合成 DNA构建体，以创建两个最常用的N.淋病 菌株MS11和FA1090。这些菌株代表细菌的不同系统发育分支。 有了这个CRISPRi的条件突变库，研究人员可以询问整个N。 淋病基因内容，以推进基础研究和疫苗或抗菌剂 发展

Abstract Neisseria gonorrhoeae is the sole causative agent of the sexually transmitted infection gonorrhea that only lives in the human population. N. gonorrhoeae has co-evolved with its human host throughout modern history and has evolved specific mechanisms of pathogenesis that relate to its mode of transmission and the anatomical sites it colonizes. Rates of gonorrhea are rising along with the rapid acquisition of antibiotic resistance. There is an urgent need to understand the mechanisms of N. gonorrhoeae pathogenesis to develop new treatment options or vaccine candidates. N. gonorrhoeae is naturally competent for DNA transformation and is one of a few bacterial species that does not regulate competence. This grant will use CRISPR interference technology (CRISPRi) to determine the essential genes of N. gonorrhoeae and provide the research community with the ability to regulate the essential genes. This work will complement our funded R24 resource grant, which creates insertional mutants of every open reading frame of N. gonorrhoeae but cannot mutate essential genes. The R24 project is leveraging the transformation competence of N. gonorrhoeae and synthetic DNA constructs to create ordered gene knockout libraries of the two most used N. gonorrhoeae strains, MS11 and FA1090. These strains represent different phylogenetic clades of the bacterium. With this conditional mutant library of CRISPRi, researchers can interrogate the entire N. gonorrhoeae gene content to advance fundamental research and vaccine or antimicrobial development.