Supplementary Figure 15: Improvement of editing specificity in the ASO:mRNA hybrid.
From: Precise RNA editing by recruiting endogenous ADARs with antisense oligonucleotides
Editing of the 5´-CAA codon to restore the E342K mutation in SERPINA1 (a reverse primer was used here!) comes along with off target editing at the nearest neighboring adenosine (see also preceding Supplementary Figure, panel B). To reduce proximal off-target editing, the 3 nt gap in the modification pattern of the ASO was reduced to a 2 nt gap by putting an additional chemical modification (2´-O-methyl uridine) opposite the off-target nucleotide. Two representative sequencing traces were selected from three very similar replicates which show that the additional chemical modification strongly reduces the proximal off-target edit while only modestly influencing the on target editing. (editing was performed in HeLa cells with SERPINA1 PiZZ cDNA overexpressed from a plasmid, 5 pmol/96 well ASO was transfected)
