Fig. 5.

Replication and immunogenicity of C/3’NCR-mir(T) virus in adult AG129 mice. a Schematic representation of viral genomes used in the study. dCGR—duplicated capsid gene region; ΔC—truncated C gene; C-opt—a full-length copy of C gene containing synonymous mutations introduced in each AA codon (except AUG and UGG); colored boxes indicate miRNA targets for mir-9–5p (cherry), mir-141–3p (purple), mir-202–5p (blue), mir-124–3p (red); 2 A—autoprotease 2 A from foot-and-mouth disease virus (FMDV); the curved arrow indicates position of 2 A protease cleavage; striped cherry and red boxes indicate mutated targets for mir-9–5p and mir-124–3p, respectively; striped green box indicates scr sequence; striped gray box indicates random sequence of 21 nt. b–h mice were infected ip with 10⁶ pfu of the indicated viruses. Mean virus titer ± SD (shown as error bars) in the serum at 1 and 3 dpi b, spleen at 3 dpi c, testis at 3 dpi d, and 12 dpi f, epididymis at 3 dpi e and 12 dpi g, and brain at 12 dpi h was determined by titration in Vero cells. The dashed lines indicate the limit of virus detection. Differences between viral titers in the mouse serum or organs at 1 or 3 dpi were compared using unpaired two-tailed t test (ns denotes not statistically significant; p > 0.05). Differences between viral titers in the organs at 12 dpi were compared using one-way ANOVA (ns p > 0.05; *p < 0.05; **p < 0.01; ****p < 0.0001). i–j Mice were infected ip with 10⁵ pfu of C/3’NCR-mir(T) and C/3’NCR-scr. At 29 days post immunization, animals were challenged with 10⁵ pfu of wt ZIKV (strain Paraiba_01/2015). Neutralizing antibody titer in the serum of immunized mice at 28 dpi i and 56 dpi (27 days post challenge) j were compared using unpaired two-tailed t test (ns denotes not statistically significant (p > 0.05]). Horizontal lines denote geometric mean ± geometric SD (shown as error bars)