Fig. 6: Transcript repair in a mouse model of Rett syndrome using circular wobble-optimized CLUSTER guide RNAs and endogenous murine Adars.

a, Editing yields in different brain regions, after delivery of the AAV-PHP.eB-encoded guide RNA through retro-orbital injection into Rett mice carrying the Mecp2 W104>amber mutation and quantification by Sanger sequencing 4 weeks later. b, Correlation between the median editing in a and the geometric mean of the RT–qPCR targets in c,g,i,k. Olfactory bulb, Ob; cerebellum, Cb; hippocampus, Hi; cortex, Cx; midbrain, Mb; thalamus, Th; brainstem, Bs. c, Guide RNA expression quantified by RT–qPCR (normalized to the geometric mean of Actb, Rps29 and Rnu6. d, As in b but correlating a,c. e, Absolute quantification of AAV episomes per cell by standard curve qPCR (normalized to Actb). f, As in b but correlating a,e. g, As in c but for Adar1. h, As in b but correlating a,g. i, As in c but for Adar1 p150. j, As in b but correlating a,i. k, As in c but for Adar2. l, As in b but correlating a,k. m, Bystander off-target events at the guide RNA-binding sites and the TS. n, All amplicon reads with on-target editing binned according to their number of bystander events (based on Supplementary Fig. 14). o, Global RNA editing at 2,533 endogenous sites (coverage ≥ 50 reads, REDIportal⁵⁵). p, Thalamus sections stained for MeCP2 and DAPI (nuclei). For a, data are shown as the median editing percentage ± 95% CI determined in n = 3 mice (nontargeting virus) and n = 5 mice (targeting virus). For statistical analysis, a Mann–Whitney U-test (two-tailed, nonparametric) was applied. For c,e,g,i,k,m,n, data are shown as the median fold change ± 95% CI or median number of copies per cell ± 95% CI determined in n = 2 mice (three technical replicates each). The NGS analysis in o is based on results from n = 1 (nontargeting virus) or n = 2 (targeting virus) mice. The results in p are derived from n = 1 mouse per group. For b,d,f,h,j,l, the R² values were determined by simple linear regression.