利用 SARS-CoV-2 刺突蛋白介导的膜融合检测法监测活细胞中病毒和靶细胞膜的融合动力学。
Monitoring fusion kinetics of viral and target cell membranes in living cells using a SARS-CoV-2 spike-protein-mediated membrane fusion assay.
机构信息
Division of Molecular Virology and Genetics, Joint Research Center for Human Retrovirus Infection, Kumamoto University, Kumamoto 8600811, Japan; Department of Clinical Pathology, Faculty of Medicine, Suez Canal University, Ismailia 41511, Egypt.
Division of Molecular Virology and Genetics, Joint Research Center for Human Retrovirus Infection, Kumamoto University, Kumamoto 8600811, Japan; Graduate School of Medical Sciences, Kumamoto University, Kumamoto 8600811, Japan.
出版信息
STAR Protoc. 2022 Sep 26;3(4):101773. doi: 10.1016/j.xpro.2022.101773. eCollection 2022 Dec 16.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein mediates membrane fusion between the virus and the target cells, triggering viral entry into the latter. Here, we describe a SARS-CoV-2 spike-protein-mediated membrane fusion assay using a dual functional split reporter protein to quantitatively monitor the fusion kinetics of the viral and target cell membranes in living cells. This approach can be applied in various cell types, potentially predicting the pathogenicity of newly emerging variants. For complete details on the use and execution of this protocol, please refer to Kimura et al. (2022b), Kimura et al. (2022c), Motozono et al. (2021), Saito et al. (2022a), Saito et al. (2022b), Suzuki et al. (2022), and Yamasoba et al. (2022).
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）刺突蛋白介导病毒与靶细胞之间的膜融合，触发病毒进入后者。在这里，我们描述了一种使用双功能分裂报告蛋白的 SARS-CoV-2 刺突蛋白介导的膜融合测定法，用于定量监测活细胞中病毒和靶细胞膜的融合动力学。该方法可应用于各种细胞类型，可能预测新出现变异体的致病性。有关该方案使用和执行的完整详细信息，请参阅 Kimura 等人（2022b）、Kimura 等人（2022c）、Motozono 等人（2021）、Saito 等人（2022a）、Saito 等人（2022b）、铃木等人（2022）和 Yamasoba 等人（2022）。
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