Fig. 3.

AAVDJ-CRISPR-CasRx confers prophylactic protection in a mouse model for hand, foot and mouth disease. a. BALB/c mice were injected intraperitoneally with a dose of 1 × 10¹¹ vgs or 1 × 10¹² vgs of AAVDJ-EV71gRNAs per mouse at 2 days old and subsequently injected intraperitoneally with EV-A71 at a dose of 2 × 10⁷ PFU per mouse at 5 days old. 1 × 10¹² vgs AAVDJ-GFPgRNA was used as control. b. Survival curve of treated mice over 19 days post-infection (dpi). c. The body weight of each mouse in each treatment group over 19 dpi. Comparison between two groups (CasRx-GFPg2 and CasRx-EV71_3Dguides) was analyzed by Log-rank (Mantel–Cox) test. ∗p < 0.05, ∗∗p < 0.005. d. The clinical score of mice was recorded using the mice clinical assessment scoring system (M-CASS) involving observation of five markers: activity, breathing, movement, body weight, and dehydration over 19 dpi. e. Treatment reduces virus titers in the hind limbs of mice as determined using viral plaque assay. Viral titration results were from a single experiment with multiple mice. f. Treatment reduces virus titers in the brain of mice as determined using viral plaque assay. Comparison between two groups (CasRx-GFPg2 and CasRx-EV71_3Dguides) was analyzed by Kruskal–Wallis test with Dunn's multiple comparisons post hoc test. ∗∗p < 0.005. Viral titration results were from a single experiment with multiple mice. g. Immuno-histochemistry staining for the EV-A71 VP2-specific antigen. Presence of viral antigen in the hind limbs and cervical spinal cord anterior horn cells (red arrow). h. H&E staining of the hindlimbs and spinal cord of mice. Polymorphonuclear meningitis in the spinal cord (black arrow). Necrosis and focal interstitial mononuclear cell infiltrates in the hind limbs and spinal cord (blue arrow). Magnification of H&E and IHC images are taken at 200X.