Genome-wide CRISPR screen identifies host factors that regulate SARS-CoV-2 entry

 

Yunkai Zhu, 

Fei Feng, 

Gaowei Hu, 

Yuyan Wang, 

Yin Yu, 

Yuanfei Zhu, 

Wei Xu, 

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 

DOIhttps://doi.org/10.1038/s41467-021-21213-4

PDF: https://www.nature.com/articles/s41467-021-21213-4.pdf

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.

https://www.nature.com/articles/s41467-021-21213-4