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. 2021 Jan 15;7:618651. doi: 10.3389/fcvm.2020.618651

Figure 1.

Figure 1

Scheme of VSV-ΔG*SARS-CoV-2 spike generation and its use in cells of interest in a 1-week protocol. On Day1, 2 transfections and 1 lentiviral infection are simultaneously performed. 293T cells are transfected with pCAGGS-G (1a), 293T cells are transfected with pCMV-3X-Flag-SARS-CoV-2 Spike (1b), and cells of interest (eg Vero-E6 cells) are infected with Lenti-ACE2, -TMPRSS2 and -Furin with PGE2 and P407 as adjuvants (1c). On Day 2, the 293T cells from 1a are inoculated with rVSVΔG-GFP*G to create an amplified working (2), and the 293T cells from 1b are inoculated with rVSVΔG-GFP*G from (2) or with a previously generated working stock of rVSVΔG-GFP*G to generate the pseudotyped rVSVΔG-GFP*SARS-CoV-2 virus (3). On Day 3 or 4, pseudotyped rVSVΔG-GFP*SARS-CoV-2 particles are collected and inoculated into the cells from 1c. On Day 5, scan the plate for GFP signal and finally collect cells for RNA/protein work (5). PGE2: Prostaglandin E2. P407: Polaxamer 407.