Figure 5 | Scientific Reports

Figure 5

From: A selectable all-in-one CRISPR prime editing piggyBac transposon allows for highly efficient gene editing in human cell lines

Figure 5

PB-PE gene editing of SOD1_R115G (A) Sanger chromatograms of D8.9 hiPSC harboring a mono-allelic SOD1_R115G point mutation after transfection of PB-PE vectors and 5 days of puromycin selection. Two different vectors were employed, one with a 15 nt RTt (PB-PE SOD1 g6 R15P15) and one with a 20 nt RTt (PB-PE SOD1 g6 R20P15). Unguided PB-PE vector served as control (PB-PE ugc). (B) Chromatograms of seven sub-clones directly established from PB-PE SOD1 g6 R15P15 treated cells. Fully gene corrected clones are denoted in blue font. (C) Colony counts of PB-PE edited D8.9 hiPSC clones and a gene edited bulk population after hyPBase_exo transfection and FIAU selection. Excised clones as determined by PB-PE qRT-PCR are shown in light green, non-excised clones are shown in light red. (D) Chromatograms of the four excised and PB-PE edited sub-clones established from a PB-PE SOD1 g6 R20P15 treated bulk population. Sub-clones with fully correct SOD1 sequences are denoted in blue font. Blue arrow: additional mutation, likely introduced by 'read-through' into pegRNA scaffold.

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