Details

Autor(en) / Beteiligte

• Chan, Charles W F
• Wang, Bei
• Nan, Lang
• Huang, Xiner
• Mao, Tianjiao
• Chu, Hoi Yee
• Luo, Cuiting
• Chu, Hin
• Choi, Gigi C G
• Shum, Ho Cheung
• Wong, Alan S L

Titel

High-throughput screening of genetic and cellular drivers of syncytium formation induced by the spike protein of SARS-CoV-2

Ist Teil von

• Nature biomedical engineering, 2024-03, Vol.8 (3), p.291-309

Ort / Verlag

England

Erscheinungsjahr

2024

Quelle

MEDLINE

Beschreibungen/Notizen

• Mapping mutations and discovering cellular determinants that cause the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to induce infected cells to form syncytia would facilitate the development of strategies for blocking the formation of such cell-cell fusion. Here we describe high-throughput screening methods based on droplet microfluidics and the size-exclusion selection of syncytia, coupled with large-scale mutagenesis and genome-wide knockout screening via clustered regularly interspaced short palindromic repeats (CRISPR), for the large-scale identification of determinants of cell-cell fusion. We used the methods to perform deep mutational scans in spike-presenting cells to pinpoint mutable syncytium-enhancing substitutions in two regions of the spike protein (the fusion peptide proximal region and the furin-cleavage site). We also used a genome-wide CRISPR screen in cells expressing the receptor angiotensin-converting enzyme 2 to identify inhibitors of clathrin-mediated endocytosis that impede syncytium formation, which we validated in hamsters infected with SARS-CoV-2. Finding genetic and cellular determinants of the formation of syncytia may reveal insights into the physiological and pathological consequences of cell-cell fusion.