Extended Data Fig. 4: AZD7648 treatment improves gene targeting at HBB, CFTR and HBA1 loci in PSC.
a. Schematic for gene targeting at the HBB locus to edit SCD (E6V) mutation in exon 1 using RNP/AAV6 gene editing (upper panel). Allelic distribution of WT, INDEL and HDR frequencies following gene editing at the HBB locus with and without AZD7648 treatment as measured by ICE analysis (lower panel) (n = 3). Mean HDR to INDEL ratio is represented above the bars. b. Schematic for gene targeting at the CFTR locus to edit CF disease mutation (ΔF508) in exon 11 using RNP/AAV6 gene editing (upper panel). Allelic distribution of WT, INDEL and HDR frequencies following gene editing with and without AZD7648 treatment at the CFTR locus as measured by ICE analysis (lower panel) (n = 3). Mean HDR to INDEL ratio is represented above the bars. c. Schematic for gene targeting at the HBB locus for knock-in of UBC-GFP-bGHpA sequence using RNP/AAV6 gene editing (upper panel). Allelic distribution of WT, INDEL and HDR frequencies following gene editing at the HBB locus with and without AZD7648 and M3814 (2 and 0.5 μM) treatment (lower panel) (n = 3). HDR efficiency was measured through ddPCR analysis. WT and INDEL frequencies were measured by ICE analysis. Mean HDR to INDEL ratio is represented above the bars. d. Schematic for gene replacement at the HBA1 locus for knock-in of transgene-2A-YFP sequence using RNP/AAV6 gene editing (upper panel). Allelic distribution of WT, INDEL and HDR frequencies following gene editing at the HBA1 locus with and without AZD7648 and M3814 (2 and 0.5 μM) treatment (lower panel) (n = 3). HDR efficiency was measured through ddPCR analysis. WT and INDEL frequencies were measured by ICE analysis. Mean HDR to INDEL ratio is represented above the bars. All data in a, b, c and d are shown as mean ± SEM.
