A CRISPRi screen of essential genes of Neisseria gonorrhoeae

摘要 淋球菌是性传播感染淋病的唯一病原体 只生活在人类群体中。淋病奈瑟菌自始至终都与人类宿主共同进化。 现代历史，并演变出与其发病模式有关的特定发病机制 传播和它殖民的解剖位置。淋病的发病率随着疾病的迅速发展而上升 获得抗生素耐药性。迫切需要了解N。 淋病发病机制研究开发新的治疗方案或候选疫苗。淋病奈瑟菌 具有天然的DNA转化能力，是少数几种不具备DNA转化能力的细菌之一 规范能力。这笔赠款将使用CRISPR干扰技术(CRISPRi)来确定 淋球菌的基本基因，并为研究界提供了调控的能力 必要的基因。这项工作将补充我们资助的R24资源赠款，这将创建 淋球菌每个开放阅读框的插入突变体，但不能突变为必需的 基因。R24项目正在利用淋球菌和合成酶的转化能力。 构建DNA载体以创建两种最常用淋病奈瑟菌的有序基因敲除文库 菌株MS11和FA1090。这些菌株代表了该细菌的不同系统发育分支。 有了CRISPRi的这个条件突变文库，研究人员可以询问整个N. 淋病基因含量促进基础研究和疫苗或抗菌药物 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.