@article{219f6177afe44e80b671c8393b98c1e7,
title = "A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion",
abstract = "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. Molecular Mechanism of Behavior; Medical Microbiology; Methodology in Biological Sciences; Cell Biology; Host-pathogen Interactions; Molecular Microbiology; Genetic Engineering; Genetic Screens",
keywords = "Cell Biology, Genetic Engineering, Genetic Screens, Host-pathogen Interactions, Medical Microbiology, Methodology in Biological Sciences, Molecular Mechanism of Behavior, Molecular Microbiology",
author = "Park, {Joseph S.} and Helble, {Jennifer D.} and Lazarus, {Jacob E.} and Guanhua Yang and Blondel, {Carlos J.} and Doench, {John G.} and Starnbach, {Michael N.} and Waldor, {Matthew K.}",
note = "Funding Information: We thank Brigid Davis and Waldor laboratory members for providing helpful comments on the project and manuscript. We are grateful to Monty Krieger, Ken Fields, Sean Whelan, and Lindsey Robinson for reagents and advice. We thank Chad Araneo for his technical assistance with FACS. J.S.P. HHMI Medical Research Fellowship; J.E.L. T32 AI-007061; C.J.B. FONDECYT 11160901, REDI170269, and an HHMI-Gulbenkian International Research Scholar; M.N.S. AI39558, AI062827, and AI113187; M.K.W. AI-042347 and HHMI. J.S.P. J.D.H. J.E.L. M.N.S. and M.K.W. conceived of the project and designed the experiments. C.J.B. and J.G.D. prepared the CRISPR library, and J.S.P. and G.Y. performed the screen, sequencing, and analysis. J.S.P. J.D.H. and J.E.L. performed the experiments. J.S.P. J.D.H. J.E.L. and M.K.W. wrote the manuscript, and all authors edited the paper. The authors declare no competing interests. Funding Information: We thank Brigid Davis and Waldor laboratory members for providing helpful comments on the project and manuscript. We are grateful to Monty Krieger, Ken Fields, Sean Whelan, and Lindsey Robinson for reagents and advice. We thank Chad Araneo for his technical assistance with FACS. J.S.P., HHMI Medical Research Fellowship; J.E.L., T32 AI-007061; C.J.B., FONDECYT 11160901 , REDI170269 , and an HHMI-Gulbenkian International Research Scholar; M.N.S., AI39558 , AI062827 , and AI113187 ; M.K.W., AI-042347 and HHMI . Publisher Copyright: {\textcopyright} 2018 The Author(s)",
year = "2019",
month = jan,
day = "25",
doi = "10.1016/j.isci.2018.12.011",
language = "English",
volume = "11",
pages = "71--84",
journal = "iScience",
issn = "2589-0042",
publisher = "Elsevier Inc.",
}