High-throughput screening of genetic and cellular drivers of syncytium formation induced by the spike protein of SARS-CoV-2.
In: Nature biomedical engineering, Jg. 8 (2024-03-01), Heft 3, S. 291-309
Online
academicJournal
Zugriff:
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.
(© 2023. The Author(s).)
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High-throughput screening of genetic and cellular drivers of syncytium formation induced by the spike protein of SARS-CoV-2.
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Autor/in / Beteiligte Person: | Chan, CWF ; Wang, B ; Nan, L ; Huang, X ; Mao, T ; Chu, HY ; Luo, C ; Chu, H ; Choi, GCG ; Shum, HC ; Wong, ASL |
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Zeitschrift: | Nature biomedical engineering, Jg. 8 (2024-03-01), Heft 3, S. 291-309 |
Veröffentlichung: | London : Springer Nature ; <i>Original Publication</i>: [London] : Macmillan Publishers Limited, [2016]-, 2024 |
Medientyp: | academicJournal |
ISSN: | 2157-846X (electronic) |
DOI: | 10.1038/s41551-023-01140-z |
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