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Nat Genet. 2017 Feb;49(2):193-203. doi: 10.1038/ng.3741. Epub 2016 Dec 19.

A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors.

Author information

1
Ragon Institute of Massachusetts General Hospital (MGH), Massachusetts Institute of Technology (MIT), and Harvard University, Cambridge, Massachusetts, USA.
2
Harvard-MIT Health Sciences and Technology, Harvard Medical School, Boston, Massachusetts, USA.
3
Broad Institute of MIT and Harvard University, Cambridge, Massachusetts, USA.
4
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
5
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA.
6
David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, Massachusetts, USA.
7
Department of Cellular and Molecular Pharmacology, California Institute for Quantitative Biosciences, QB3, University of California at San Francisco (UCSF), San Francisco, California, USA.
8
Gladstone Institute of Virology and Immunology, J. David Gladstone Institutes, San Francisco, California, USA.
9
Biological Sciences in Public Health, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA.
10
Howard Hughes Medical Institute, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
11
Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, California, USA.
12
Diabetes Center, University of California at San Francisco, San Francisco, California, USA.
13
Department of Medicine, University of California at San Francisco, San Francisco, California, USA.
14
Innovative Genomics Initiative (IGI), University of California, Berkeley, Berkeley, California, USA.
15
UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, California, USA.
16
Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
17
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.
18
Massachusetts General Hospital Cancer Center, Boston, Massachusetts, USA.
19
Institute of Medical Engineering and Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

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.

PMID:
27992415
PMCID:
PMC5511375
DOI:
10.1038/ng.3741
[Indexed for MEDLINE]
Free PMC Article

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