Figure 2.

Activity of the CRISPR-Cas9 Targets in the TLR3 System

(A) Guide RNAs are cloned into the px459 vector under the hU6 promoter. (B) px459 Cas9 targets were expressed in the HEK-TLR3 cell line. Seventy-two hours after transfection, flow cytometric analysis displayed about 27% of BFP⁺ (NHEJ repair) cells by px-459-T7. The graphs represent triplicate data from three independent experiments. Statistical analysis: ANOVA, ***p < 0.001. (C) Analysis of sequence modifications after T3 cleavage. Sequence changes were analyzed using Sanger sequencing via TOPO Cloning. Sequence in red: PAM site; sequence in blue: guide RNA target sequence; sequence in yellow: changes to the wild-type sequence. T0, px459 without guide RNA; T3, target 3.