A CRISPRi screen of essential genes of Neisseria gonorrhoeae

淋病奈瑟菌必需基因的 CRISPRi 筛选

基本信息

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
项目类别：
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

项目摘要

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.

抽象的淋病奈瑟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与其人类宿主共同进化现代历史并已进化出与其模式相关的特定发病机制传播及其定殖的解剖部位。淋病发病率随着疾病的快速增长而上升获得抗生素耐药性。迫切需要了解 N 的机制。淋病发病机制以开发新的治疗方案或候选疫苗。淋病奈瑟菌天然具有 DNA 转化能力，并且是少数不具有 DNA 转化能力的细菌物种之一规范能力。这笔赠款将使用 CRISPR 干扰技术 (CRISPRi) 来确定淋病奈瑟菌的必需基因，并为研究界提供调节的能力必需的基因。这项工作将补充我们资助的 R24 资源赠款，该赠款创建淋病奈瑟菌每个开放阅读框的插入突变体，但不能发生必需突变基因。 R24 项目正在利用淋病奈瑟菌的转化能力和合成 DNA 构建体用于创建两种最常用的淋病奈瑟菌的有序基因敲除文库菌株 MS11 和 FA1090。这些菌株代表了细菌的不同系统发育分支。借助这个 CRISPRi 条件突变文库，研究人员可以询问整个 N. 淋病基因内容可促进基础研究和疫苗或抗菌药物发展。