A genome-wide CRISPR screen identifies host factors that regulate SARS-CoV-2 entry.

Affiliation

Zhu Y(#)(1), Feng F(#)(1), Hu G(#)(1), Wang Y(#)(1), Yu Y(1), Zhu Y(1), Xu W(1), Cai X(1), Sun Z(1), Han W(1), Ye R(1), Qu D(1), Ding Q(2), Huang X(3), Chen H(4), Xu W(5), Xie Y(1), Cai Q(6), Yuan Z(7), Zhang R(8).
Author information:
(1)Key Laboratory of Medical Molecular Virology
(MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Biosafety Level 3 Laboratory, Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, China.
(2)Center for Infectious Disease Research, School of Medicine, Tsinghua University, Beijing, China.
(3)Technical Center For Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, Shanghai, China.
(4)Shanghai Veterinary Research Institute, CAAS, Shanghai, China.
(5)Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China.
(6)Key Laboratory of Medical Molecular Virology
(MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Biosafety Level 3 Laboratory, Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, China. [Email]
(7)Key Laboratory of Medical Molecular Virology
(MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Biosafety Level 3 Laboratory, Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, China. [Email]
(8)Key Laboratory of Medical Molecular Virology
(MOE/NHC/CAMS), School of Basic Medical Sciences, Shanghai Medical College, Biosafety Level 3 Laboratory, Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, China. [Email]
(#)Contributed equally

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.