Figure 2.

Light-dependent genome editing. (A) Experimental workflow. (B, C) HEK293T cells were co-transduced with AAV vectors (B) or co-transfected with plasmids (C) encoding (i) NmeCas9 and a sgRNA targeting the indicated locus and (ii) the indicated Acr variant. Cells were then irradiated with pulsed blue light or kept in the dark for 72 hours, followed by assessment of indel frequencies using NGS (B) or TIDE sequencing (C). Plasmid mass ratios used during the transfection in C are indicated. (D) Huh7 cells were co-transduced with AAV vectors encoding (i) NmeCas9 and a sgRNA targeting the indicated locus and (ii) the respective Acr. Seventy-two hours post-transduction, editing frequencies were determined by TIDE sequencing. (B-D) Bars represent mean values, error bars the standard deviation and dots individual data points from n = 3 independent experiments.