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

Resultado de la investigación: Article

Resumen

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

Idioma originalEnglish
Páginas (desde-hasta)71-84
Número de páginas14
PublicacióniScience
Volumen11
DOI
EstadoPublished - 25 ene 2019

Huella dactilar

Clustered Regularly Interspaced Short Palindromic Repeats
Chlamydia trachomatis
Heparitin Sulfate
Flow Cytometry
Genome
Microbiology
Host-Pathogen Interactions
Genetic Engineering
Biological Science Disciplines
Libraries
Cell Biology
Epithelial Cells
Bacteria
Genes

ASJC Scopus subject areas

  • General

Citar esto

Park, Joseph S. ; Helble, Jennifer D. ; Lazarus, Jacob E. ; Yang, Guanhua ; Blondel, Carlos J. ; Doench, John G. ; Starnbach, Michael N. ; Waldor, Matthew K. / A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion. En: iScience. 2019 ; Vol. 11. pp. 71-84.
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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",
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Park, JS, Helble, JD, Lazarus, JE, Yang, G, Blondel, CJ, Doench, JG, Starnbach, MN & Waldor, MK 2019, 'A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion', iScience, vol. 11, pp. 71-84. https://doi.org/10.1016/j.isci.2018.12.011

A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion. / Park, Joseph S.; Helble, Jennifer D.; Lazarus, Jacob E.; Yang, Guanhua; Blondel, Carlos J.; Doench, John G.; Starnbach, Michael N.; Waldor, Matthew K.

En: iScience, Vol. 11, 25.01.2019, p. 71-84.

Resultado de la investigación: Article

TY - JOUR

T1 - A FACS-Based Genome-wide CRISPR Screen Reveals a Requirement for COPI in Chlamydia trachomatis Invasion

AU - Park, Joseph S.

AU - Helble, Jennifer D.

AU - Lazarus, Jacob E.

AU - Yang, Guanhua

AU - Blondel, Carlos J.

AU - Doench, John G.

AU - Starnbach, Michael N.

AU - Waldor, Matthew K.

PY - 2019/1/25

Y1 - 2019/1/25

N2 - 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

AB - 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

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KW - Genetic Engineering

KW - Genetic Screens

KW - Host-pathogen Interactions

KW - Medical Microbiology

KW - Methodology in Biological Sciences

KW - Molecular Mechanism of Behavior

KW - Molecular Microbiology

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M3 - Article

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EP - 84

JO - iScience

JF - iScience

SN - 2589-0042

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