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. 2021 Sep 9;95(19):e00461-21. doi: 10.1128/JVI.00461-21

FIG 2.

FIG 2

CasRx preventing viral infection. (A) Expression of CP after transfection with CasRx-NLS and single-target CP2 (CP2-crRNA) or RdRp2 (RdRp2-crRNA) crRNAs or combined-target CP2 and RdRp2 crRNA (mix-crRNA). (B) Expression of RdRp after transfection with CasRx-NLS and CP2-crRNA, RdRp2-crRNA or mix-crRNA. (C) Expression of CP after transfection with CasRx-dNLS and CP2-crRNA, RdRp2-crRNA or mix-crRNA. (D) Expression of RdRp after transfection with CasRx-dNLS and CP2-crRNA, RdRp2-crRNA or mix-crRNA. (E and F) Cell morphology and CPEs (vacuolation) after infection with RGNNV, with or without CasRx-NLS (E) or CasRx-dNLS (F). Scale bar = 20 μm. (G) Schematic representation of the CasRx system preventing RGNNV infection at different time points. (H to K) The CasRx system prevented RGNNV infection at different time points when transfected with either CasRx-dNLS (H and I) or CasRx-NLS (J and K) and crRNAs targeting the RGNNV CP gene (CP2), RdRp gene (RdRp2), or both (mix). (H) Expression of CP after RGNNV infection 12 h, 24 h, 48 h at cells transfected with CasRx-dNLS and crRNAs targeting the RGNNV CP gene (CP2), RdRp gene (RdRp2), or both (mix). (I) Expression of RdRp after RGNNV infection 12 h, 24 h, 48 h at cells transfected with CasRx-dNLS and crRNAs targeting the CP2, RdRp2, or mix. (J) Expression of CP after RGNNV infection 12 h, 24 h, 48 h at cells transfected with CasRx-NLS and crRNAs targeting the CP2, RdRp2, or mix. (K) Expression of RdRp after RGNNV infection 12 h, 24 h, 48 h at cells transfected with CasRx-NLS and crRNAs targeting the CP2, RdRp2, or mix. (L and M) Cell morphology and CPEs (vacuolation) at different time points after RGNVV infection in cells treated with CasRx-NLS (L) or CasRx-dNLS (M). Arrows indicate the vacuoles induced by RGNNV infection. Scale bar = 20 μm. (N) Schematic representation of the CasRx system preventing RGNNV infection at different crRNA concentrations. (O) Expression of CP after transfection with CasRx-dNLS and different concentrations (50, 100, 200, 400, or 800 ng) of crRNAs (CP2, RdRp2, or mix). (P) Expression of RdRp after transfection with CasRx-dNLS and different concentrations (50, 100, 200, 400, or 800 ng) of crRNAs (CP2, RdRp2, or mix). (Q) Expression of CP after transfection with CasRx-NLS and different concentrations (50, 100, 200, 400, or 800 ng) of crRNAs (CP2, RdRp2, or mix). (R) Expression of RdRp after transfection with CasRx-NLS and different concentrations (50, 100, 200, 400, or 800 ng) of crRNAs (CP2, RdRp2, or mix). Data are expressed as mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.