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[Preprint]. 2020 Nov 2:2020.10.28.359042. Originally published 2020 Oct 28. [Version 2] doi: 10.1101/2020.10.28.359042

Fig. 1. Design and demonstration of a GFP-based activity reporter of SARS-CoV-2 main protease Mpro.

Fig. 1.

(a) Schematic of the reporter. (b) Sequence of the flipped GFP10–11. (c) Construct of the reporter FlipGFPMpro. (d) Fluorescence images (left) and quantitative analysis (right) of SARS-CoV-2 or mock-infected HEK293T cells that co-expressed hACE2. The images in the FlipGFP channel were brightened 30-fold compared to those in (e). (e) Fluorescence images of HEK293T cells expressing FlipGFPMpro and mCherry, together with the inactive Mpro mutant C145A (upper panels) or wild type Mpro (lower panels). (f) Normalized FlipGFP fluorescence by mCherry. The ratio of FlipGFP/mCherry for the Mpro/C145A is normalized to 1. Data are mean ± SD (n = 5). FlipGFPTEV is a TEV activity reporter containing TEV cleavage sequence in FlipGFP. Scale bar: 5 μm (d); 10 μm (f).