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

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 overexpr...

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Main Authors: Ravi K Dinesh, Chengkun Wang, Yuanhao Qu, Arjun Rustagi, Henry Cousins, James Zengel, Xiaotong Wang, Trisha R Barnard, William A Johnson, Guangxue Xu, Tianyi Zhang, Nicholas Magazine, Aimee Beck, Lucas Miecho Heilbroner, Grace Peters-Schulze, Aaron J Wilk, Mengdi Wang, Weishan Huang, Brooke E Howitt, Jan Carette, Russ Altman, Catherine A Blish, Le Cong
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-07-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1013157
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Summary: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.
ISSN:1553-7366
1553-7374