活细胞中新型冠状病毒M蛋白功能获得性抑制试验
Gain-of-function assay for SARS-CoV-2 M inhibition in living cells.
作者信息
Moghadasi Seyad Arad, Becker Jordan T, Belica Christopher, Wick Chloe, Brown William L, Harris Reuben S
机构信息
Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA, 55455.
Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA, 55455.
出版信息
bioRxiv. 2020 Nov 9:2020.11.09.375139. doi: 10.1101/2020.11.09.375139.
The main protease, M , of SARS-CoV-2 is required to cleave the viral polyprotein into precise functional units for virus replication and pathogenesis. Here we demonstrate a quantitative reporter for M function in living cells, in which protease inhibition by genetic or chemical methods results in strong eGFP fluorescence. This robust gain-of-function system readily distinguishes between inhibitor potencies and can be scaled-up to high-throughput platforms for drug testing.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的主要蛋白酶Mpro是将病毒多聚蛋白切割成精确功能单元以实现病毒复制和致病所必需的。在此，我们展示了一种用于活细胞中Mpro功能的定量报告系统，其中通过基因或化学方法抑制蛋白酶会导致强烈的增强型绿色荧光蛋白（eGFP）荧光。这种强大的功能获得系统能够轻松区分抑制剂的效力，并且可以扩展到高通量平台进行药物测试。
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