Fig. 3. Coupling of exonuclease to Cas9 via CC increases the length of genomic deletions.

HEK293 cells (2*10⁵ cells/ml) were transfected with plasmids encoding target gRNA and CCExo. Genomic DNA was isolated 48 h post transfection, the targeted region PCR amplified and subjected to NGS analysis. NGS analysis was performed for 200 bp amplicons for gRNA targeting MYD88 gene (a) or 400 bp amplicons for gRNA targeting CD47 gene (b), VEGFα gene (c) or EMX1 gene (d). Diagrams are showing percentage (calculated based on number of total mutated reads) of deletions (Δ) of various lengths. Genome wide determinations of potential off-target activities of CCExo for VEGFα sites in HEK293 cells and for TRBC1 site in human K562 cells, predicted by CIRCLE-seq (e).