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

PLoS Pathog. 2025 Jul 17;21(7):e1013157. doi: 10.1371/journal.ppat.1013157.

Ravi K Dinesh ¹, Chengkun Wang ¹ ², Yuanhao Qu ¹ ³, Arjun Rustagi ⁴ ⁵, Henry Cousins ¹ ⁶, James Zengel ⁷, Xiaotong Wang ¹, Trisha R Barnard ⁴, William A Johnson ¹, Guangxue Xu ¹, Tianyi Zhang ⁷, Nicholas Magazine ⁸, Aimee Beck ⁴, Lucas Miecho Heilbroner ¹, Grace Peters-Schulze ¹, Aaron J Wilk ⁴ ⁶, Mengdi Wang ⁹, Weishan Huang ⁸ ¹⁰, Brooke E Howitt ¹, Jan Carette ⁶, Russ Altman ² ¹¹, Catherine A Blish ⁴ ⁶ ¹², Le Cong ¹ ² ³

Affiliations

1 Department of Pathology, Stanford University School of Medicine, Stanford, California, United States of America.
2 Department of Genetics, Stanford University School of Medicine, Stanford, California, United States of America.
3 Cancer Biology Program, Stanford University School of Medicine, Stanford, California, United States of America.
4 Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, United States of America.
5 Department of Medicine, University of California, San Francisco (UCSF), San Francisco, California, United States of America.
6 Medical Scientist Training Program, Stanford University School of Medicine, Stanford, California, United States of America.
7 Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America.
8 Department of Pathology, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, United States of America.
9 Department of Electrical and Computer Engineering, Center for Statistics and Machine Learning, Princeton University, Princeton, New Jersey, United States of America.
10 Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, New York, United States of America.
11 Department of Bioengineering, Stanford University, Stanford, California, United States of America.
12 Chan Zuckerberg Biohub, San Francisco, California, United States of America.

PMID: 40674406 DOI: 10.1371/journal.ppat.1013157

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.

Products

Cat. No.

Product Name

Description

Target

Research Area
HY-112205A

RR-11a analog

99.12%, Legumain Inhibitor

Parasite; Legumain

Infection

Name
Email *

	Sorry, but the email address you supplied was invalid.