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. 2020 Nov 4;6(45):eabb2154. doi: 10.1126/sciadv.abb2154

Fig. 1. Generation of ZIKV vaccines using SMARRT.

Fig. 1

(A) Schematic of the ZIKV prM-E and ZIKV NS3 vaccines. All structural genes were removed from the Venezuelan equine encephalitis virus (strain TC-83) while retaining the four nonstructural proteins (nsP1 to nsP4) that encode the replicase. prM-E or NS3 genes from ZIKV strain SPH2015 replaced the structural genes downstream of the viral 26S promoter. BHK-21 cells were electroporated with water (Mock), an irrelevant RNA (bridging control), or RNA encoding either ZIKV prM-E or NS3 and then analyzed 20 hours after electroporation. UTR, untranslated region. (B) Western blot analysis of whole-cell lysates probed with anti-ZIKV E or NS3 Abs. MW, molecular weight. (C) Quantification of launch efficiency by intracellular staining of BHK-21 cells with an anti-dsRNA (J2) Ab. Data are presented as the mean ± SD of triplicates from one experiment, representative of two independent experiments. The nonparametric Mann-Whitney test was used to compare Mock versus each group; ****P < 0.0001.