A CRISPRi screen of essential genes of Neisseria gonorrhoeae

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.