Fig. 1: Rational attenuation of SARS-CoV-2 WA1/2020. | Nature Communications

Fig. 1: Rational attenuation of SARS-CoV-2 WA1/2020.

From: Intranasal delivery of a rationally attenuated SARS-CoV-2 is immunogenic and protective in Syrian hamsters

Fig. 1

a Top, genome organization of SARS-CoV-2. Leader sequence (red), transcriptional regulatory sequence within the leader sequence (TRS-L) and within the body (TRS-B) are highlighted in green. Bottom, either the polybasic insert “PRRA” (red) alone or together with ORF6-8 (green) were removed from the WA1/2020 genome. Locations of K164A/H165A and N128S/K129E are indicated at the bottom left of the figure. b Representative images of plaques formed by individual recombinant virus in Vero E6 cells. c Sanger sequencing result of Nsp1-K164A/H165A virus after passage 5. d A549-hACE2 cells were inoculated with indicated virus (n = 3 biological replicates) at multiplicity of infection (MOI) of 0.01. Output virus in the supernatants were determined on Vero E6 cells by plaque assay. e MatTek EpiAirway cells in 24-well plates were inoculated with indicated virus at MOI of 2. Supernatants collected at 2 (n = 6), 24 (n = 6), 48 (n = 5), 72 (n = 4), 96 (n = 3), and 120 (n = 2) hours post-infection (HPI) were titrated using a focus-forming assays biological replicates. Error bars for d and e indicate standard deviation. Data presented are representatives of two independent experiments with multiple biological replicates. Source data are provided as a Source data file. f MatTek EpiAirway cells from (e) were fixed in 10% formalin followed by immunostaining of nucleocapsid protein (in green) at 1, 2, 3, 5 dpi with one biological replicate per time point. Nuclei were counterstained by DAPI (in blue). Scale bar, 300 μm.

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