Extended Data Fig. 8: AAV9 dosage effect on IHCs in mice.
From: Template-independent genome editing and restoration for correcting frameshift disorders
a, Whole mount immunostaining of WT cochleae demonstrating 400 nl AAV9-CAG-mCherry (1×1013 GC/ml) induces mCherry expression in 100% of IHCs. mCherry (red), Myo7a (yellow), and DAPI (blue) signals are shown. Right panels highlight magnified regions (white frames) from the left panels. Scale bars: 200 μm (left), 20μm (right). Multiple mice were tested to validate this observation. b, Whole mount immunostaining of Otof1233delC/1233delC cochleae revealing dual AAV9(SpCas9 + gRNA1) dosage-dependent Otoferlin expression in IHCs. Otoferlin (green), Myo7a (red), and DAPI (blue) signals are shown. The injected dual AAV9 volume was from 400, 600, 800, 1000, to 1500 nl, as indicated on the top of panels. Scale bars: 100 μm. c, Quantification of Otoferlin expressing IHC in uninjected and dual AAV9 injected Otof1233delC/1233delC mice. d, Quantification of click and pure-tone ABR thresholds from graded volume of dual AAV9 treated Otof1233delC/1233delC mice (green), in comparison with WT mice (gray) and untreated Otof1233delC/1233delC mice (purple).
