Fig. 4: In vivo base editing in NHPs.
From: High-efficiency base editing in the retina in primates and human tissues
a, Schematic of the SABE(N) and SABE(C) constructs. b, Dual AAV5-SABE vectors were delivered to 23 eyes of NHPs by subretinal injection under the macula. Illustration in a, illustrations of the AAV ITRs and AAV capsid in a and illustration of the eye in b created with BioRender.com. Some of the injections also detached the fovea. OCT was used to confirm successful bleb formation. c, Experimental design. The number of eyes with fovea detached are indicated in brackets. d, Representative immunofluorescence images of ABE(N) expression in a NHP retinal section (scale bars, 20 μm). IS/OS, photoreceptor inner and outer segments; ONL, outer nuclear layer (photoreceptors); OPL, outer plexiform layer; INL, inner nuclear layer. Gray, Hoechst; cyan, ABE(N); magenta, arrestin3; gamma correction has been applied to obtain an optimal dynamic range for visualization. e, In vivo base-editing efficiencies in gDNA and ABCA4 mRNA in the retina with different base-editor constructs and at different doses. Results are presented as the mean ± s.d. Significance for dose response was calculated using a one-way ANOVA with Tukey’s correction, (gDNA: *P = 0.011, ABCA4 mRNA: *P = 0.011). f, In vivo base-editing efficiencies in gDNA and ABCA4 mRNA in the RPE/choroid with different base-editor constructs and at different doses. Results are presented as the mean ± s.d. g, In vivo base-editing efficiencies in gDNA of sorted cones and rods and representative immunofluorescence images of sorted cells (scale bars, 25 μm). IS, photoreceptor inner segment. Results are presented as the mean ± s.d. White, Hoechst; magenta, arrestin3; orange, rhodopsin. h, In vivo base-editing efficiencies in the foveal retina and RPE/choroid in gDNA at different dose levels and with different constructs. Results are presented as the mean ± s.d.
