Transduction of AAV Vectors in Subretinally Injected Mice
(A) Transduction profile of AAV vectors following subretinal injection (SRI) in Nrl-GFP mice. AAV-mediated mCherry expression in representative fluorescent fundus images (top row) and retinal cross-sections (middle and bottom rows) taken at 4 weeks post-injection with 2e12 vg/mL is shown. (B) Transduction profile of AAV44.9(E531D)-IRBP/GNAT2-GFP following SRI in C57BL/6J mice. Six weeks post-injection with 1 μL of 2e12 vg/mL (2e9 vg delivered), retinal cross-sections were stained with an antibody directed against cone arrestin and counterstained with DAPI. Vector-mediated GFP expression and cone arrestin colocalize, indicating that this capsid efficiently transduces cone photoreceptors. (C) Transduction efficiencies as determined by flow cytometry in subretinally injected Nrl-GFP mice. ∗Kruskal-Wallis tests were performed, followed by a post hoc Dunn’s test to make pairwise comparisons between groups. For the non-rod values, H = 44.912, df = 6, and p < 0.001. For the rod values, H = 85.546, df = 6, and p < 0.001. Scale bars in (A) and (B), 50 μm. RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer.