A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion

• Published in: iScience Vol. 11; pp. 71 - 84
• Main Authors: Park, Joseph S., Helble, Jennifer D., Lazarus, Jacob E., Yang, Guanhua, Blondel, Carlos J., Doench, John G., Starnbach, Michael N., Waldor, Matthew K.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25.01.2019; Elsevier
• Subjects: Cell Biology; Genetic Engineering; Genetic Screens; Host-pathogen Interactions; Medical Microbiology; Methodology in Biological Sciences; Molecular Mechanism of Behavior; Molecular Microbiology; Medical Microbiology; Host-pathogen Interactions; Genetic Engineering; Molecular Mechanism of Behavior; Genetic Screens; Methodology in Biological Sciences; Cell Biology; Molecular Microbiology
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2018.12.011

The invasion of Chlamydia trachomatis, an obligate intracellular bacterium, into epithelial cells is driven by a complex interplay of host and bacterial factors. To comprehensively define the host genes required for pathogen invasion, we undertook a fluorescence-activated cell sorting (FACS)-based CRISPR screen in human cells. A genome-wide loss-of-function library was infected with fluorescent C. trachomatis and then sorted to enrich for invasion-deficient mutants. The screen identified heparan sulfate, a known pathogen receptor, as well as coatomer complex I (COPI). We found that COPI, through a previously unappreciated role, promotes heparan sulfate cell surface presentation, thereby facilitating C. trachomatis attachment. The heparan sulfate defect does not fully account for the resistance of COPI mutants. COPI also promotes the activity of the pathogen's type III secretion system. Together, our findings establish the requirement for COPI in C. trachomatis invasion and the utility of FACS-based CRISPR screening for the elucidation of host factors required for pathogen invasion. [Display omitted] •FACS-based CRISPR screen to identify host factors required for C. trachomatis invasion•Candidate genes comprise heparan sulfate biosynthesis, actin remodeling, and COPI•COPI regulates heparan sulfate cell surface presentation and C. trachomatis attachment•COPI is also required for efficient C. trachomatis T3SS translocation Molecular Mechanism of Behavior; Medical Microbiology; Methodology in Biological Sciences; Cell Biology; Host-pathogen Interactions; Molecular Microbiology; Genetic Engineering; Genetic Screens