Figure 2
From: CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
CRISPR Del/Rei of rs12293670 (NRGN) in the RU02 iPSC line. (a) sgRNA, syn-sgRNA, and ssODN repair template design. (b-e) Sanger sequencing chromatograms of the cell line before and after each step of editing. (f) Screen of the bulk cells from each deletion transfection well. Each lane represents a technical replicate from a separate transfection well unless otherwise marked. Editing efficiency quantified by Image J shown under each lane. (g) Example deletion clone screening gel. Each lane represents one clone. Lanes marked with a red cross were positive clones confirmed by Sanger sequencing. (h) Screen of the bulk cells from the reinsertion transfection. (i) Example screen of reinsertion clones derived from single-cell cloning. (j) Summary of total clones screened for each step. The positive by electrophoresis percentage refers to the number of clones that looked positive by gel out of the total clones amplified. The confirmed by sequencing percentage refers to the number of clones that were confirmed positive by Sanger sequencing out of the total number of clones sent for sequencing. Photographs of original gels available in Supplemental Fig. 6. All ladders are 1 Kb plus. UE = unedited/reinsertion positive control, NTC = no template negative control.
