A genome-wide CRISPR screen identifies host factors that regulate SARS-CoV-2 entry
Top Cited Papers
Open Access
- 11 February 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 12 (1), 1-11
- https://doi.org/10.1038/s41467-021-21213-4
Abstract
The global spread of SARS-CoV-2 is posing major public health challenges. One feature of SARS-CoV-2 spike protein is the insertion of multi-basic residues at the S1/S2 subunit cleavage site. Here, we find that the virus with intact spike (Sfull) preferentially enters cells via fusion at the plasma membrane, whereas a clone (Sdel) with deletion disrupting the multi-basic S1/S2 site utilizes an endosomal entry pathway. Using Sdel as model, we perform a genome-wide CRISPR screen and identify several endosomal entry-specific regulators. Experimental validation of hits from the CRISPR screen shows that host factors regulating the surface expression of angiotensin-converting enzyme 2 (ACE2) affect entry of Sfull virus. Animal-to-animal transmission with the Sdel virus is reduced compared to Sfull in the hamster model. These findings highlight the critical role of the S1/S2 boundary of SARS-CoV-2 spike protein in modulating virus entry and transmission and provide insights into entry of coronaviruses.Keywords
Funding Information
- the Natural Science Foundation of Shanghai
- Project of Novel Coronavirus Research of Fudan University
- Development Programs for COVID-19 of Shanghai Science and Technology Commission
- National Natural Science Foundation of China (32041005)
This publication has 62 references indexed in Scilit:
- Retromer: A Master Conductor of Endosome SortingCold Spring Harbor Perspectives in Biology, 2014
- Genome-Scale CRISPR-Cas9 Knockout Screening in Human CellsScience, 2014
- Propagation, Quantification, Detection, and Storage of West Nile VirusCurrent Protocols in Microbiology, 2013
- A Short Hairpin RNA Screen of Interferon-Stimulated Genes Identifies a Novel Negative Regulator of the Cellular Antiviral ResponsemBio, 2013
- Development of a Highly Protective Combination Monoclonal Antibody Therapy against Chikungunya VirusPLoS Pathogens, 2013
- Simultaneous Treatment of Human Bronchial Epithelial Cells with Serine and Cysteine Protease Inhibitors Prevents Severe Acute Respiratory Syndrome Coronavirus EntryJournal of Virology, 2012
- Assembly and Solution Structure of the Core Retromer Protein ComplexTraffic, 2010
- Cholesterol Synthesis Inhibitor U18666A and the Role of Sterol Metabolism and Trafficking in Numerous Pathophysiological ProcessesLipids, 2009
- Inhibitors of cathepsin L prevent severe acute respiratory syndrome coronavirus entryProceedings of the National Academy of Sciences of the United States of America, 2005
- COMMD Proteins, a Novel Family of Structural and Functional Homologs of MURR1Journal of Biological Chemistry, 2005