Extended Data Fig. 2: Optimization of in vivo administration route.
From: In vivo base editing extends lifespan of a humanized mouse model of prion disease
a, Experimental design for assessment of in vivo administration route. 4-week-old Tg66 mice were treated with dual-AAV PHP.eB BE3.9max for installation of PRNP R37X by systemic administration through retro-orbital injection or direct CNS administration through intracerebroventricular (ICV) stereotaxic injection. Mice treated with retro-orbital injection received a total of 4x1012 vg/kg AAV (2x1012 vg/kg each for N- and C-terminal AAV) (n=2). Mice treated with ICV injection received a total of 5.5x1011 vg/kg AAV (2.5x1011 vg/kg each for N- and C-terminal AAV and 5.0x1010 vg/kg for AAV encoding EGFP fused to nuclear membrane-localized Klarsicht/ANC-1/Syne-1 homology (KASH) domain (EGFP:KASH)) (n=2). Mouse brains were harvested five weeks after treatment. GFP-positive nuclei were sorted from ICV-injected samples via FACS to enrich for AAV-transduced cells. Genomic DNA extracted from the brain tissues were analyzed for editing efficiency. b, Frequency of the desired R37X edit in the bulk brain hemisphere of mice untreated (n=2) or treated with dual-AAV PHP.eB BE3.9max by retro-orbital injection (n=2), or by ICV injection (n=2). Dots represent individual biological replicates and bar graphs represent mean values.
