Ordered gene knockout libraries for Neisseria gonorrhoeae

有序的淋病奈瑟菌基因敲除文库

基本信息

批准号：
10649487
负责人：
Hank S. SEIFERT
金额：
$ 61.71万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10649487
关键词：
Anatomy Antibiotic Resistance Backcrossings Bacteria Bar Codes California Case Study Codon Nucleotides Communities Competence Consensus DNA Development Disease Ensure Gene Order Genes Genetic Genetic Recombination Genome Gonorrhea Grant Human Individual Infection International Investigation Kanamycin Resistance Knowledge Laboratories Letters Libraries Link Modern 1601-history Mutate Mutation National Institute of Allergy and Infectious Disease Neisseria Neisseria gonorrhoeae Open Reading Frames Organism Pathogenesis Pathogenicity Persons Phylogenetic Analysis Physiology Population Reporting Research Research Personnel Resource Sharing Resources Schedule Sexually Transmitted Agents Sexually Transmitted Diseases Suggestion Uncertainty Vaccines Work design design and construction drug-resistant gonorrhea genetic resource human pathogen innovation knockout gene meetings mutant pathogen programs resistance gene resistance mechanism screening 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 is restricted to the human population. N. gonorrhoeae has co-evolved with its human host throughout all 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 a rapid acquisition of antibiotic resistance. There is an urgent need to understand of 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 resource grant will leverage the transformation competence of N. gonorrhoeae and synthetic DNA constructs to create ordered gene knockout libraries of the two most commonly used N. gonorrhoeae strains, MS11 and FA1090. These strains represent different phylogenetic clades of the bacterium. The libraries will include deletions of all genes encoding an open reading frame (ORF) that will allow the replacement of the ORF with a non-polar kanamycin resistance gene (KmR). Each KmR insertion mutant will also carry a unique random barcode. Leading researchers in the field and NIAID program officials also support this resource proposal as an important shared resource to help advance progress in this field.

抽象的淋病奈瑟菌是性传播感染淋病的唯一病原体，仅限于人类。淋病奈瑟菌自始至终与其人类宿主共同进化整个现代历史，并已进化出与其发病模式相关的特定发病机制。传播及其定殖的解剖部位。淋病发病率正在迅速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。该资源补助将利用 N 的转化能力。淋病和合成 DNA 构建体，以创建两种最常见的有序基因敲除文库常用的淋病奈瑟菌菌株为MS11和FA1090。这些菌株代表了不同的细菌的系统发育枝。该文库将包括所有编码基因的删除开放阅读框 (ORF)，允许用非极性卡那霉素替换 ORF 抗性基因（KmR）。每个 KmR 插入突变体还将携带一个独特的随机条形码。该领域的领先研究人员和 NIAID 项目官员也支持这一资源提案，因为帮助推动这一领域进步的重要共享资源。