Correction of m.4291T > C in primary patient-derived fibroblasts. Top left: The primary patient-derived fibroblast line is homoplasmic for the m.4291T > C mutation. Top right: Design of two sets of Left (red) and Right (green) target sequences flanking the spacer region (blue) for TALE-guided editing in MT-TI to correct m.4291C > T (dark red). Mismatched T (red) in TALE target sequence L2. Middle: Overview of the transfection strategy of DdCBE for primary skin fibroblasts. Bottom left: Editing efficiencies of all four combinations Left- and Right-TALE-DdCBE plasmids in primary patient-derived fibroblasts as measured by amplicon sequencing (Illumina NGS). Bottom middle: Ratio of on-target editing vs. bystander editing at any other base within the spacing region in fibroblast cells. Bottom right: Clonal lines of edited primary patient-derived fibroblasts show high variability in mitochondrial heteroplasmy indicating different editing efficiencies per cell (Illumina NGS).
What is the therapeutic potential of correcting mutations by base editing of the #MitochondrialGenome? This study shows that #mitochondrial #BaseEditing can indeed functionally create & correct mitochondrial pathogenic #mutations in patient-derived cells @plosbiology.org 🧪 plos.io/3ZMa2K2