A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion
| Autor: | Jacob E. Lazarus, Guanhua Yang, Carlos J. Blondel, Joseph Park, Jennifer D. Helble, John G. Doench, Michael N. Starnbach, Matthew K. Waldor |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Genetic Screens 02 engineering and technology Biology medicine.disease_cause Article Type three secretion system 03 medical and health sciences chemistry.chemical_compound medicine CRISPR lcsh:Science Pathogen Multidisciplinary Molecular Microbiology Cell Biology COPI Heparan sulfate Cell sorting 021001 nanoscience & nanotechnology 3. Good health Cell biology 030104 developmental biology chemistry Medical Microbiology Methodology in Biological Sciences Host-pathogen Interactions lcsh:Q Molecular Mechanism of Behavior Genetic Engineering 0210 nano-technology Chlamydia trachomatis Genetic screen |
| Zdroj: | iScience, Vol 11, Iss, Pp 71-84 (2019) iScience |
| ISSN: | 2589-0042 |
| Popis: | Summary 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. Graphical Abstract Highlights • 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 |
| Databáze: | OpenAIRE |
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