A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion
Joseph S. Park,
Jennifer D. Helble,
Jacob E. Lazarus,
Guanhua Yang,
Carlos J. Blondel,
John G. Doench,
Michael N. Starnbach,
Matthew K. Waldor
Affiliations
Joseph S. Park
Howard Hughes Medical Institute, Boston, MA 02215, USA; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA; Division of Infectious Diseases, Brigham & Women's Hospital, Boston 02115, MA, USA; Boston University School of Medicine, Boston, MA 02120, USA
Jennifer D. Helble
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA
Jacob E. Lazarus
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA; Division of Infectious Diseases, Brigham & Women's Hospital, Boston 02115, MA, USA; Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA 02114, USA
Guanhua Yang
Division of Infectious Diseases, Brigham & Women's Hospital, Boston 02115, MA, USA
Carlos J. Blondel
Division of Infectious Diseases, Brigham & Women's Hospital, Boston 02115, MA, USA
John G. Doench
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
Michael N. Starnbach
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA
Matthew K. Waldor
Howard Hughes Medical Institute, Boston, MA 02215, USA; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA; Division of Infectious Diseases, Brigham & Women's Hospital, Boston 02115, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Corresponding author
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. : Molecular Mechanism of Behavior; Medical Microbiology; Methodology in Biological Sciences; Cell Biology; Host-pathogen Interactions; Molecular Microbiology; Genetic Engineering; Genetic Screens Subject Areas: Molecular Mechanism of Behavior, Medical Microbiology, Methodology in Biological Sciences, Cell Biology, Host-pathogen Interactions, Molecular Microbiology, Genetic Engineering, Genetic Screens
