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Fig.10. Off-target analysis after in vitro and in vivo base editing with HDAdEF1a.ABE8e (based on in silico prediction). A) Analysis based on in silico OTS prediction using Cas-OFFinder software for the mouse genome. 74 sites were nominated, from which the top 5 were subjected to NGS. Amplicons were prepared using genomic DNA template from a naïve mouse and an in vivo transduced mouse with the highest on-targeting editing rate. The middle and lower panels show the frequency of A>G conversions and indels, respectively. B) Analysis based on in silico OTS prediction using Cas-OFFinder software for the human genome. 89 sites were nominated, from which the top 5 were subjected to NGS. Amplicons were prepared using genomic DNA from erythroid cells derived from CD34+ cells of a thalassemia patient that were in vitro transduced with HDAd-EF1a.ABE8e.