A Genome-wide CRISPR Screen Identifies Host Factors That Regulate SARS-CoV-2 Entry

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Feb 12

PMID 33574281

Citations 150

Authors

Yunkai Zhu

Fei Feng

Gaowei Hu

Yuyan Wang

Yin Yu

Yuanfei Zhu

Xia Cai

Zhiping Sun

Wendong Han

Rong Ye

Di Qu

Qiang Ding

Xinxin Huang

Hongjun Chen

Wei Xu

Youhua Xie

Qiliang Cai

Zhenghong Yuan

Rong Zhang

Affiliations

Soon will be listed here.

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.

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