Extended Data Fig. 4. Incorporation of epegRNAs and PEmax together substantially enhance prime editing in CNS.

a-b, PE3-AAVs encoding a Dnmt1 +1 C-to-G edit were delivered via P0 ICV injection as v1, v1e, or v1em architecture at a dose of 1x10¹¹ vg (5x10¹⁰ vg each N- and C-terminal AAVs) with 1x10¹⁰ vg promoter-matched AAV9 EGFP:KASH. At three weeks, nuclei from cortex were isolated and sorted by FACS to either all nuclei (bulk) or GFP+ populations. Genomic DNA was extracted and analyzed by HTS. While improvement from incorporation of epegRNA alone did not reach statistical significance (P=0.17), incorporation of epegRNA and PEmax (v1em) yielded highly statistically significant improvement in prime editing over v1 (for both bulk and GFP+ populations, P<0.0001). Significance was calculated by two-tailed unpaired t-test with correction for multiple comparisons. Dots represent individual data points and error bars represent mean±SEM for n = 3 different mice.