Fig. 1: Construction of MyoAAV-UA and evaluation of its efficiency in activating utrophin in two Dmd mouse models.
a Schematic diagram of the dCasMINI-VPR construct used for utrophin activation. b–d RT-qPCR analysis showing high efficiency of combined sgRNAs in activating utrophin mRNA in Neuro 2a cell line (b), cynomolgus monkey skeletal muscle cells (MSkM) (c), and HEK293T cell line (d). Data are normalized to control (mock-sgRNA). n = 3 technical replicates in each group. e Schematic of the MyoAAV-UA expression cassette and its application in Dmd mice, NHPs, and humans. f Representative immunoblots of dystrophin and utrophin in quadriceps muscles from WT group, mdx mice treated with saline, mdx mice treated with MyoAAV-UA, DmdΔEx50h mice treated with saline, and DmdΔEx50 mice treated with MyoAAV-UA. Vinculin was used as a loading control. g Representative immunofluorescence staining of dystrophin (green), utrophin (red), and DAPI (blue) in quadriceps muscles from different groups. Scale bar, 100 µm. h Quantification of utrophin protein levels in quadriceps from different groups were normalized to vinculin (n = 6 mice in each group). i Quantification of utrophin mRNA expression levels in quadriceps muscles from different groups, normalized to Gapdh (n = 6 mice in each group). f, g The experiment was repeated with six biologically independent replicates with similar results. h, i Data are normalized to WT (n = 6 biologically independent experiments in each group). b–d, h, i Data are presented as mean ± s.d. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. Source data are provided as a Source Data file.
