Fig. 2: FiCAT precision characterization.
From: Find and cut-and-transfer (FiCAT) mammalian genome engineering
a Characterization of junctions generated by targeted insertion at the ITR. Cells were transfected with FiCAT R372A_K375A_D450N, ½ GFP transposon and TRAC gRNA. The number of reads containing deletions (black) and insertions (blue) produced at the ITR site are plotted by position from the start of the ITR. Representative sequencing reads are shown at the bottom. Reference target sequence and ITR annotation are highlighted, the gRNA cutting site is indicated with a vertical black line and deletions and insertions are shown in black and blue, respectively. b Characterization of insertions generated by FiCAT R372A_K375A_D450N mutant (top panel) and Cas9-PB WT sample (bottom panel) in Hek293T cell line transfected with an RFP transposon and TRAC gRNA in cell population enriched by flow cytometry in RFP positive cells 16 days after transfection. Significant insertion frequencies are shown in vertical lines (FDR q-value < 0.001). On-target insertions on the TRAC loci are shown in green and off-target insertions in red.
