Extended Data Fig. 4: Assessment of on-target and off-target editing by guide RNAs targeting the HPD gene.
From: Improved specificity and efficiency of in vivo adenine base editing therapies with hybrid guide RNAs
a, Lentiviral screening in primary human hepatocytes for adenine base editing of heterologous HPD splice site target sequences by various gRNAs in combination with ABE8.20; target adenines are in bold and underlined. MIT score = off-target/specificity score indicated by UCSC Genome Browser. Four candidate gRNAs (HPD3, HPD4, HPD5, and HPD20) were taken forward for off-target analyses. b, Targeted amplicon sequencing of ONE-seq-nominated sites in plasmid-treated vs. control wild-type HuH-7 cells with HPD3, HPD4, HPD5, or HPD20 gRNA in combination with ABE8.8 (n = 2 treated and 2 untreated biological replicates). Sites with unsuccessful sequencing are omitted. Arrows indicate verified sites of off-target editing. Means are shown.
