Membrane-wide screening identifies potential tissue-specific determinants of SARS-CoV-2 tropism

Authors

• Ravi K. Dinesh, Stanford University School of Medicine, Stanford, CA
• Chengkun Wang, Stanford University School of Medicine, Stanford, CA
• Yuanhao Qu, Stanford University School of Medicine, Stanford, CA
• Arjun Rustagi, Stanford University School of Medicine, Stanford, CA
• Henry Cousins, Stanford University School of Medicine, Stanford, CA
• James Zengel, Stanford University School of Medicine, Stanford, CA
• Xiaotong Wang, Stanford University School of Medicine, Stanford, CA
• Trisha R. Barnard, Stanford University School of Medicine, Stanford, CA
• William A. Johnson, Stanford University School of Medicine, Stanford, CA
• Guangxue Xu, Stanford University School of Medicine, Stanford, CA
• Tianyi Zhang, Stanford University School of Medicine, Stanford, CA
• Nicholas Magazine, Louisiana State University School of Veterinary Medicine, Baton Rouge, LA
• Aimee Beck, Stanford University School of Medicine, Stanford, CA
• Lucas Miecho Heilbroner, Stanford University School of Medicine, Stanford, CA
• Grace Peters-Schulze, Stanford University School of Medicine, Stanford, CA
• Aaron J. Wilk, Stanford University School of Medicine, Stanford, CA
• Mengdi Wang, Princeton University, Princeton, NJ
• Weishan Huang, Louisiana State University School of Veterinary Medicine, Baton Rouge, LA
• Brooke E. Howitt, Stanford University School of Medicine, Stanford, CA
• Jan Carette, Stanford University School of Medicine, Stanford, CA
• Russ Altman, Stanford University School of Medicine, Stanford, CA
• Catherine A. Blish, Stanford University School of Medicine, Stanford, CA
• Le Cong, Stanford University School of Medicine, Stanford, CA

Document Type

Article

Publication Date

7-17-2025

Publication Title

PLoS Pathogens

Abstract

While SARS-CoV-2 primarily infects the respiratory tract, clinical evidence indicates that cells from diverse cell types and organs are also susceptible to infection. Using the CRISPR activation (CRISPRa) approach, we systematically targeted human membrane proteins in cells with and without overexpression of ACE2, thus identifying unrecognized host factors that may facilitate viral entry. Validation experiments with replication-competent SARS-CoV-2 confirmed the role of newly identified host factors, particularly the endo-lysosomal protease legumain (LGMN) and the potassium channel KCNA6, upon exogenous overexpression. In orthogonal experiments, we show that disruption of endogenous LGMN or KCNA6 decreases viral infection and that inhibitors of candidate factors can reduce viral entry. Additionally, using clinical data, we find possible associations between expression of either LGMN or KCNA6 and SARS-CoV-2 infection in human tissues. Our results identify potentially druggable host factors involved in SARS-CoV-2 entry, and demonstrate the utility of focused, membrane-wide CRISPRa screens in uncovering tissue-specific entry factors of emerging pathogens.

First Page

1

Last Page

29

DOI

10.1371/journal.ppat.1013157

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Dinesh, Ravi K.; Wang, Chengkun; Qu, Yuanhao; Rustagi, Arjun; Cousins, Henry; Zengel, James; Wang, Xiaotong; Barnard, Trisha R.; Johnson, William A.; Xu, Guangxue; Zhang, Tianyi; Magazine, Nicholas; Beck, Aimee; Heilbroner, Lucas Miecho; Peters-Schulze, Grace; Wilk, Aaron J.; Wang, Mengdi; Huang, Weishan; Howitt, Brooke E.; Carette, Jan; Altman, Russ; Blish, Catherine A.; and Cong, Le, "Membrane-wide screening identifies potential tissue-specific determinants of SARS-CoV-2 tropism" (2025). LSU-LCMC Cancer Center Faculty Publications. 197.
https://digitalscholar.lsuhsc.edu/llcc_facpubs/197
10.1371/journal.ppat.1013157