A CRISPRi screen of essential genes of Neisseria gonorrhoeae

淋病奈瑟菌必需基因的 CRISPRi 筛选

• 批准号: 10656956
• 负责人: Hank S. SEIFERT
• 金额: $ 24万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-21 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656956
• 关键词: Anatomy; Antibiotic Resistance; Bacteria; CRISPR interference; CRISPR library; California; Case Study; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Competence; Complement; Consensus; DNA; Development; Disease; Essential Genes; Funding; Gene Expression Regulation; Gene Order; Genes; Genetic; Gonorrhea; Grant; Human; Individual; Infection; International; Investigation; Knock-out; Knowledge; Laboratories; Letters; Libraries; Michigan; Modern 1601-history; Mutate; Neisseria; Neisseria gonorrhoeae; Neisseria lactamica; Neisseria meningitidis; Open Reading Frames; Organism; Pathogenesis; Pathogenicity; Phylogenetic Analysis; Physiology; Plasmids; Population; R24; Reporting; Research; Research Personnel; Resource Sharing; Resources; Sexually Transmitted Agents; Sexually Transmitted Diseases; Suggestion; System; Technology; Uncertainty; Universities; Vaccines; Work; antimicrobial; conditional mutant; drug-resistant gonorrhea; fundamental research; genetic resource; human pathogen; innovation; knockout gene; mutant; pathogen; resistance mechanism; symposium; synthetic construct; transmission process; vaccine candidate

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.

抽象的 Neisseria Gonorrhoeae是性传播感染淋病的唯一病因 只生活在人口中。 N. Gonorrhoeae与其人类宿主共同发展 现代历史，并发展了与其模式有关的发病机理的特定机制 传输及其定居的解剖位点。淋病的发生率随着快速的速度而上升 获得抗生素耐药性。迫切需要了解N的机制。 淋病的发病机理开发新的治疗选择或候选疫苗。 N. Gonorrhoeae 自然具有DNA转化的能力，并且是几种不符合DNA的细菌物种之一 规范能力。该赠款将使用CRISPR干扰技术（CRISPRI）确定 N.淋病的基本基因，并为研究界提供了调节的能力 基本基因。这项工作将补充我们资助的R24资源赠款 淋病猪笼草的每个开放阅读框架的插入突变体，但不能突变必需 基因。 R24项目正在利用N.淋病和合成的转化能力 DNA构建体以创建有序的基因敲除库的两个最常用的淋病猪笼草的库 菌株，MS11和FA1090。这些菌株代表了细菌的不同系统发育进化枝。 有了这个条件的CRISPRI突变库，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素 发展。