Supplementary Figure 6: TIDE sequencing analysis of light-mediated indel mutation.
From: Engineered anti-CRISPR proteins for optogenetic control of CRISPR–Cas9
HEK293T cells were co-transduced with AAV vectors expressing the Cas9, CASANOVA and a gRNA targeting the CCR5 (a) or EMX1 (b) locus. Cells were exposed to blue light for 70 h or kept in the dark as control, followed by TIDE sequencing1. The target locus was PCR-amplified with primers flanking the expected breakpoint followed by Sanger sequencing of the amplicon. Total editing efficiencies and frequencies of individual insertions or deletions were then calculated by decomposition of the sequencing chromatogram using the TIDE web tool (https://www.deskgen.com/landing/tide.html). TIDE sequencing revealed a broad range of different insertions and deletions in the light, but not in the dark control samples. Data represent a single experiment.
