FIGURE 3.

Successful treatment in murine PKU following ex‐vivo gene therapy. (A) Change of blood Phe levels in male (n = 3) and female (n = 3) PKU mice during paracetamol dosing. The dashed lines represent the therapeutic threshold of 360 μM and “normal” blood Phe level, which is below 120 μM, and the thick black lines represent the mean Phe level (Locally Weighted Scatterplot Smoothing using R). Note that one dose of paracetamol was given per week. At the end of the experiments, mice were sacrificed to determine PAH enzyme activity (B) and gene editing efficiency (C) in the resected livers. The % of PAH enzyme activity is presented relative to wild‐type from all treated mice (the dashed line represents the PAH mean activity of PKU mice). Gene editing in individual mice was quantified by Cypor indel frequency determined by next generation sequencing (NGS). In addition, the percentage of wild‐type Pah copy number was determined by NGS, and all mice were transplanted with 2000 vg/cell treated primary hepatocytes unless indicated otherwise (° = 1000 vg/cell). Statistics: (A) two‐way ANOVA for unbalanced designs, (B) and (C) one‐way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.