Fig. 1: In vivo correction of Pahenu2 mice using AAV-mediated delivery of intein-split PEmax.
From: Treatment of a metabolic liver disease in mice with a transient prime editing approach
a–c, Correction (top) and indel rates (bottom) in HEK293T cells with the integrated Pahenu2 locus, transfected with PE2 versus PEmax (n = 5) (a), with unmodified (grey) versus tevopreQ1 (tevo) modified versus tmpknot (tmp) modified pegRNAs (Supplementary Table 1) (n = 4) (b) or with tevopreQ1-mPKU-2.1 versus tevopreQ1-mPKU-SM (n = 4) (c). Values represent mean ± s.d. of independent biological replicates. Means were compared using unpaired two-sided Student’s t-tests. NS, not significant, P > 0.05. d, Schematic illustration of the experimental outline of the analysis in adult PKU mice. e, Schematic illustration of the AAV constructs selected for in vivo experiments. Indicated are AAV genome lengths including inverted terminal repeats (ITRs) in base pairs (bp). W3, truncated version of the posttranscriptional regulatory element of woodchuck hepatitis virus; nSpCas9, nickase of Streptococcus pyogenes Cas9; tevo, trimmed engineered pegRNA; M-MLV-RTdRnH, M-MLV RT-delta RNase H; SV40-pA, simian virus 40 poly-adenylation signal; NLS, nuclear localization signal. f, Correction rates in PKU mice (n = 3 mice). Editing rates were assessed in lysed tail tissue, liver tissue and isolated hepatocytes. Values depict mean ± s.d. Hep, hepatocytes. g, Phe levels at the experimental end point of untreated homozygous Pahenu2 mice (UT −/−) (n = 5), untreated heterozygous mice (UT +/−) (n = 3) and homozygous treated mice (n = 3). Dotted lines indicate recommended therapeutic thresholds for Phe levels in adults (600 μmol l−1) or in children/during pregnancy (360 μmol l−1)29,57. Values represent mean ± s.d. of independent biological replicates and were analysed using an ordinary one-way analysis of variance using Dunnett’s multiple comparisons test.
