Evaluation of off-target profile by transcriptome-wide analyses in vitro

We analyzed the effect of bystander A-to-G editing and found no introduction of premature STOP mutations indicating therapeutic potential of xABE in treating DMD caused by c.4174C>T. To evaluate transcriptome-wide off-target effect of xABE targeting c.4174C>T, we profiled different xABE driven by EFS (E1), CBh (C3, C4) and MHCK7 (M2) promoter with a non-targeting xABE as control by RNA-seq. Our results found non-significant difference of A-to-G/C-to-U SNV number generated by E1/C3/C4/M2 and control xABE, suggesting high editing specificity of xABE targeting c.4174C>T at transcriptome scale.