title: A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors.
abstract: Host proteins are essential for HIV entry and replication and can be important nonviral therapeutic targets. Large-scale RNA interference (RNAi)-based screens have identified nearly a thousand candidate host factors, but there is little agreement among studies and few factors have been validated. Here we demonstrate that a genome-wide CRISPR-based screen identifies host factors in a physiologically relevant cell system. We identify five factors, including the HIV co-receptors CD4 and CCR5, that are required for HIV infection yet are dispensable for cellular proliferation and viability. Tyrosylprotein sulfotransferase 2 (TPST2) and solute carrier family 35 member B2 (SLC35B2) function in a common pathway to sulfate CCR5 on extracellular tyrosine residues, facilitating CCR5 recognition by the HIV envelope. Activated leukocyte cell adhesion molecule (ALCAM) mediates cell aggregation, which is required for cell-to-cell HIV transmission. We validated these pathways in primary human CD4+T cells through Cas9-mediated knockout and antibody blockade. Our findings indicate that HIV infection and replication rely on a limited set of host-dispensable genes and suggest that these pathways can be studied for therapeutic intervention.
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.|Harvard-MIT Health Sciences and Technology, Harvard Medical School, Boston, Massachusetts, USA.|Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Park
authors.affiliation: Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.|Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.|David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA.
authors.forename: Tim
authors.initials: T
authors.lastname: Wang
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Dylan
authors.initials: D
authors.lastname: Koundakjian
authors.affiliation: Department of Cellular and Molecular Pharmacology, California Institute for Quantitative Biosciences, QB3, University of California at San Francisco (UCSF), San Francisco, California, USA.|Gladstone Institute of Virology and Immunology, J. David Gladstone Institutes, San Francisco, California, USA.
authors.forename: Judd F
authors.initials: JF
authors.lastname: Hultquist
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.|Biological Sciences in Public Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA.
authors.forename: Pedro
authors.initials: P
authors.lastname: Lamothe-Molina
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.|Howard Hughes Medical Institute, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Blandine
authors.initials: B
authors.lastname: Monel
authors.affiliation: Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, California, USA.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Schumann
authors.affiliation: Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Yu
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Krupzcak
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.|Harvard-MIT Health Sciences and Technology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Wilfredo
authors.initials: W
authors.lastname: Garcia-Beltran
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.
authors.forename: Alicja
authors.initials: A
authors.lastname: Piechocka-Trocha
authors.affiliation: Department of Cellular and Molecular Pharmacology, California Institute for Quantitative Biosciences, QB3, University of California at San Francisco (UCSF), San Francisco, California, USA.|Gladstone Institute of Virology and Immunology, J. David Gladstone Institutes, San Francisco, California, USA.
authors.forename: Nevan J
authors.initials: NJ
authors.lastname: Krogan
authors.affiliation: Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, California, USA.|Diabetes Center, University of California at San Francisco, San Francisco, California, USA.|Department of Medicine, University of California at San Francisco, San Francisco, California, USA.|Innovative Genomics Initiative (IGI), University of California, Berkeley, Berkeley, California, USA.|UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Marson
authors.affiliation: Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.|Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.|David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA.|Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Sabatini
authors.affiliation: Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.|Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Lander
authors.affiliation: Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.|Massachusetts General Hospital Cancer Center, Boston, Massachusetts, USA.
authors.forename: Nir
authors.initials: N
authors.lastname: Hacohen
authors.affiliation: Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.|Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.|Howard Hughes Medical Institute, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.|Institute of Medical Engineering and Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Walker
mesh_terms: D020410:Activated-Leukocyte Cell Adhesion Molecule / Q000235:genetics; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D016678:Genome / Q000235:genetics; D015658:HIV Infections / Q000235:genetics*; D015497:HIV-1 / Q000472:pathogenicity; D054884:Host-Pathogen Interactions / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D034622:RNA Interference / Q000502:physiology; D019713:Receptors, CCR5 / Q000235:genetics; D015238:Sulfotransferases / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: D020410:Activated-Leukocyte Cell Adhesion Molecule; C584933:CCR5 protein, human; D008565:Membrane Proteins; D019713:Receptors, CCR5; D015238:Sulfotransferases; C114740:TPST2 protein, human