Fig. 5. Improved prime editing in the mouse CNS.

Comparison of prime editing from v1em and v3em PEmax AAVs in the CNS via direct injection in neonatal mice or systemic injection in adult mice. a, Percent GFP⁺ nuclei of capsid-matched, promoter-matched and terminator-matched EGFP:KASH used for enrichment. Dots represent individual mice, and error bars represent mean ± s.e.m. of n = 3 mice; each condition includes both male and female mice. b, Either v1em or v3em PE3-AAV9 encoding the +1 C-to-G edit at Dnmt1 was delivered via P0 ICV injection at a dose of 1 × 10¹¹ vg. Cortex was harvested after 3 weeks, and nuclei were isolated. GFP⁺ subpopulations were sorted by FACS. gDNA from bulk or GFP⁺ nuclei was extracted and analyzed by HTS. Dots represent individual mice, and error bars represent mean ± s.e.m. of n = 3 mice; each condition includes both male and female mice. c, Adult 6–8-week-old mice were RO injected with a total dose of 1 × 10¹² vg AAV PHP.eB with either v1em or v3em PE3-AAV encoding the +2 G-to-C edit at Dnmt1. Cortex was harvested after 3 weeks, and nuclei were isolated. GFP⁺ subpopulations were sorted by FACS. gDNA from bulk or GFP⁺ nuclei was extracted and analyzed by HTS. Dots represent individual mice, and error bars represent mean ± s.e.m. of n = 4–5 mice; each condition includes both male and female mice. d, Installation of APOE3 R136S (APOE Christchurch) in humanized APOE3 mice. 1 × 10¹¹ vg v3em PE3-AAV9 was administered by ICV injection to mice on P1 or P3. gDNA or total RNA was isolated from neocortex and hippocampus (matched hemispheres). RNA was converted to cDNA, and both gDNA and cDNA were analyzed by HTS. Dots represent individual mice, and error bars represent mean ± s.e.m. of n = 3–4 mice; each condition includes both male and female mice.