Figure 7.

DSB Repair Fidelity after Double Cutting

(A) Repair products that may result from Cas9 in combination with two sgRNAs that target adjacent sequences. All products are amplified by PCR and detected by high-throughput sequencing.

(B) Quantification of the various repair products 60 hr after Cas9 induction in the presence of sgRNA-LBR2 and one of 5 sgRNAs (labeled 5 through 9) that target a sequence within 110–300 bp from the LBR2 site.

(C) Same as (B) but highlighting the relative proportions of perfect and imperfect junctions among the repaired excision events.

(D) Relative proportions of indels at sgRNA-LBR2 target site (blue), indels at sgRNA-LBR8 target site (red), indels at both sgRNA-LBR2 and sgRNA-LBR8 target sites (green), excised DNA (yellow), and wild-type (black) fractions over time. The data points are normalized to the total mutation fraction to correct for the variation in transfection efficiency. Average of 2 independent experiments is shown; error bars represent the SD. Dashed lines show fitted sigmoid curves.