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

作者：Zhu, Yunkai; Feng, Fei; Hu, Gaowei; Wang, Yuyan; Yu, Yin; Zhu, Yuanfei; Xu, Wei; Cai, Xia; Sun, Zhiping; Han, Wendong; Ye, Rong; Qu, Di; Ding, Qiang; Huang, Xinxin; Chen, Hongjun; Xu, Wei; Xie, Youhua; Cai, Qiliang^*; Yuan, Zhenghong^*; Zhang, Rong^*

来源：Nature Communications, 2021, 12(1): 961.

DOI：10.1038/s41467-021-21213-4

• RESPIRATORY SYNDROME CORONAVIRUS
• CELL ENTRY
• INHIBITOR U18666A
• SPIKE PROTEIN
• TRAFFICKING
• RETROMER
• COMPLEX
• MECHANISMS
• TARGET
• NPC1

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. The SARS-CoV-2 spike protein contains a multi-basic cleavage site. Here, the authors show how this multi-basic cleavage site affects entry of SARS-CoV-2 into cells and transmission in the hamster model and identify host factors affecting entry of SARS-CoV-2 in a genome-wide CRISPR screen.

• 单位
  中国医学科学院; 南方医科大学; 清华大学; 复旦大学