tetano
Editor, Senior Moderator
PLoS Biol
. 2025 Jun 12;23(6):e3002738.
doi: 10.1371/journal.pbio.3002738. Online ahead of print. Global siRNA screen identifies human host factors critical for SARS-CoV-2 replication and late stages of infection
Xin Yin[SUP] 1 [/SUP], Yuan Pu[SUP] 2 [/SUP], Shuofeng Yuan[SUP] 3 [/SUP], Lars Pache[SUP] 4 [/SUP], Christopher Churas[SUP] 5 [/SUP], Stuart Weston[SUP] 6 [/SUP], Laura Riva[SUP] 7 [/SUP], Lacy M Simons[SUP] 8 [/SUP], William J Cisneros[SUP] 8 [/SUP], Thomas Clausen[SUP] 9 [/SUP], Grace Biddle[SUP] 6 [/SUP], Simon Doss-Gollin[SUP] 6 [/SUP], Meagan Deming[SUP] 6 [/SUP], Paul D De Jesus[SUP] 2 [/SUP], Ha Na Kim[SUP] 10 [/SUP], Daniel Fuentes[SUP] 2 [/SUP], John M Whitelock[SUP] 10 [/SUP], Jeffrey D Esko[SUP] 9 [/SUP], Megan S Lord[SUP] 10 [/SUP], Ignacio Mena[SUP] 2 [/SUP], Adolfo García-Sastre[SUP] 11 12 13 14 15 16 [/SUP], Judd F Hultquist[SUP] 8 [/SUP], Matthew B Frieman[SUP] 6 [/SUP], Trey Ideker[SUP] 5 17 [/SUP], Dexter Pratt[SUP] 5 [/SUP], Laura Martin-Sancho[SUP] 18 [/SUP], Sumit K Chanda[SUP] 2 7 [/SUP]
Affiliations
Defining the subset of cellular factors governing SARS-CoV-2 replication can provide critical insights into viral pathogenesis and identify targets for host-directed antiviral therapies. While a number of genetic screens have previously reported SARS-CoV-2 host dependency factors, most of these approaches relied on utilizing pooled genome-scale CRISPR libraries, which are biased toward the discovery of host proteins impacting early stages of viral replication. To identify host factors involved throughout the SARS-CoV-2 infectious cycle, we conducted an arrayed genome-scale siRNA screen. Resulting data were integrated with published functional screens and proteomics data to reveal (i) common pathways that were identified in all OMICs datasets-including regulation of Wnt signaling and gap junctions, (ii) pathways uniquely identified in this screen-including NADH oxidation, or (iii) pathways supported by this screen and proteomics data but not published functional screens-including arachionate production and MAPK signaling. The identified proviral host factors were mapped into the SARS-CoV-2 infectious cycle, including 32 proteins that were determined to impact viral replication and 27 impacting late stages of infection, respectively. Additionally, a subset of proteins was tested across other coronaviruses revealing a subset of proviral factors that were conserved across pandemic SARS-CoV-2, epidemic SARS-CoV-1 and MERS-CoV, and the seasonal coronavirus OC43-CoV. Further studies illuminated a role for the heparan sulfate proteoglycan perlecan in SARS-CoV-2 viral entry and found that inhibition of the non-canonical NF-kB pathway through targeting of BIRC2 restricts SARS-CoV-2 replication both in vitro and in vivo. These studies provide critical insight into the landscape of virus-host interactions driving SARS-CoV-2 replication as well as valuable targets for host-directed antivirals.
. 2025 Jun 12;23(6):e3002738.
doi: 10.1371/journal.pbio.3002738. Online ahead of print. Global siRNA screen identifies human host factors critical for SARS-CoV-2 replication and late stages of infection
Xin Yin[SUP] 1 [/SUP], Yuan Pu[SUP] 2 [/SUP], Shuofeng Yuan[SUP] 3 [/SUP], Lars Pache[SUP] 4 [/SUP], Christopher Churas[SUP] 5 [/SUP], Stuart Weston[SUP] 6 [/SUP], Laura Riva[SUP] 7 [/SUP], Lacy M Simons[SUP] 8 [/SUP], William J Cisneros[SUP] 8 [/SUP], Thomas Clausen[SUP] 9 [/SUP], Grace Biddle[SUP] 6 [/SUP], Simon Doss-Gollin[SUP] 6 [/SUP], Meagan Deming[SUP] 6 [/SUP], Paul D De Jesus[SUP] 2 [/SUP], Ha Na Kim[SUP] 10 [/SUP], Daniel Fuentes[SUP] 2 [/SUP], John M Whitelock[SUP] 10 [/SUP], Jeffrey D Esko[SUP] 9 [/SUP], Megan S Lord[SUP] 10 [/SUP], Ignacio Mena[SUP] 2 [/SUP], Adolfo García-Sastre[SUP] 11 12 13 14 15 16 [/SUP], Judd F Hultquist[SUP] 8 [/SUP], Matthew B Frieman[SUP] 6 [/SUP], Trey Ideker[SUP] 5 17 [/SUP], Dexter Pratt[SUP] 5 [/SUP], Laura Martin-Sancho[SUP] 18 [/SUP], Sumit K Chanda[SUP] 2 7 [/SUP]
Affiliations
- PMID: 40504864
- DOI: 10.1371/journal.pbio.3002738
Defining the subset of cellular factors governing SARS-CoV-2 replication can provide critical insights into viral pathogenesis and identify targets for host-directed antiviral therapies. While a number of genetic screens have previously reported SARS-CoV-2 host dependency factors, most of these approaches relied on utilizing pooled genome-scale CRISPR libraries, which are biased toward the discovery of host proteins impacting early stages of viral replication. To identify host factors involved throughout the SARS-CoV-2 infectious cycle, we conducted an arrayed genome-scale siRNA screen. Resulting data were integrated with published functional screens and proteomics data to reveal (i) common pathways that were identified in all OMICs datasets-including regulation of Wnt signaling and gap junctions, (ii) pathways uniquely identified in this screen-including NADH oxidation, or (iii) pathways supported by this screen and proteomics data but not published functional screens-including arachionate production and MAPK signaling. The identified proviral host factors were mapped into the SARS-CoV-2 infectious cycle, including 32 proteins that were determined to impact viral replication and 27 impacting late stages of infection, respectively. Additionally, a subset of proteins was tested across other coronaviruses revealing a subset of proviral factors that were conserved across pandemic SARS-CoV-2, epidemic SARS-CoV-1 and MERS-CoV, and the seasonal coronavirus OC43-CoV. Further studies illuminated a role for the heparan sulfate proteoglycan perlecan in SARS-CoV-2 viral entry and found that inhibition of the non-canonical NF-kB pathway through targeting of BIRC2 restricts SARS-CoV-2 replication both in vitro and in vivo. These studies provide critical insight into the landscape of virus-host interactions driving SARS-CoV-2 replication as well as valuable targets for host-directed antivirals.