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.

摘要