Skip to main content
. 2022 Jul 8;14(7):1505. doi: 10.3390/v14071505

Figure 3.

Figure 3

Translation efficiency of the SARS-CoV-2 5′UTRs. (A) Scheme of the gRNA and the different sgRNAs, with the leader sequence in black and the variable region in orange, which represent each 5′UTR (this panel was obtained from Figure 1). The sequences corresponding to each 5′UTR have been cloned upstream of the Renilla ORF. (B,C) RRL and (D) untreated RRL have been programmed with capped and polyadenylated mRNAs harboring specifically the 5′UTR of the ORF1ab, ORF-S, ORF-3a, ORF-E, ORF-M, ORF-6, ORF-7a, ORF-7b, ORF-8 or ORF-N. The resulting protein products have been resolved on (B) an SDS-PAGE and quantified with a phosphoImager, or (C,D) analyzed by luciferase assay. (E) Jurkat cells were electroporated with capped and polyadenylated mRNAs as indicated. After 90 min of incubation, the cellular extracts were used for RT-qPCR and luciferase assay. All the results are normalized to the globin 5′UTR expression (set at 100%). Values are the mean (+/− S.D.) for three independent experiments (panels (B,C)) or (+/− S.E.M.) for four independent experiments (panels (D,E)).