A genome-wide CRISPR screen identified host genes essential for intracellular Brucella survival

作者：Heling Xu, Jingjing Lu, Fang Huang, Qi Zhang, Shuang Liu, Zeliang Chen, Shanhu Li · 发表于：Microbiology Spectrum · 年份：2024 · DOI：10.1128/spectrum.03383-23 · 被引用次数：7 · 研究领域：Brucella: diagnosis, epidemiology, treatment、CRISPR and Genetic Engineering、Burkholderia infections and melioidosis

ABSTRACT Brucella is a zoonotic intracellular bacterium that poses threats to human health and economic security. Intracellular infection is a hallmark of the agent Brucella and a primary cause of distress, through which the bacterium regulates the host intracellular environment to promote its own colonization and replication, evading host immunity and pharmaceutical killing. Current studies of Brucella intracellular processes are typically premised on bacterial phenotype such as intracellular bacterial survival, followed by biochemical or molecular biological approaches to reveal detailed mechanisms. While such processes can deepen the understanding of Brucella -host interaction, the insights into host alterations in infection would be easily restricted to known pathways. In the current study, we applied CRISPR Cas9 screen to identify host genes that are most affected by Brucella infection on cell viability at the genomic level. As a result of CRISPR screening, we firstly identified that knockout of the negatively selected genes GOLGA6L6 , DEFB103B , OR4F29 , and ERCC6 attenuate the viability of both the host cells and intracellular Brucella , suggesting these genes to be potential therapeutic targets for Brucella control. In particular, knockout of DEFB103B diminished Brucella intracellular survival by altering host cell autophagy. Conversely, knockout of positive screening genes promoted intracellular proliferation of Brucella . In summary, we screened host genes at the ge...