Figure 5.

Measuring the Effect of a Therapeutic Transgene in rAAV5-Transduced Cerebral Organoids

(A) rAAV5-CAG-miATXN3 vDNA was measured by qPCR and depicted as gc per cell. rAAV5-miATXN3-transduced organoids contained on average 94 gc of vDNA/cell (rAAV5-miATXN3: n = 3 batches of 3 organoids each, Mock: n = 2 batches of 3 organoids each, rAAV5-GFP: n = 1 batch of 3 organoids; one-way ANOVA followed by Tukey’s multiple comparison, ∗∗∗p < 0.0005, ∗∗∗∗p < 0.00005; graphs represent mean and min to max values). (B) miRNA was measured in cerebral organoids by qPCR. rAAV5-miATXN3-transduced organoids expressed on average 13 miRNA copies per cell (see A; n = 3; one-way ANOVA followed by Tukey’s multiple comparison, ∗p < 0.05; graphs represent mean and min to max values). (C) Ataxin-3 protein was measured by TR-FRET in lysates of cerebral organoids transduced by rAAV5-miATXN3, or control-transduced organoids expressed 30% less ataxin-3 protein as measured by TR-FRET (see A; n = 3; one-way ANOVA followed by Tukey’s multiple comparison, ∗p < 0.05; graphs represent mean with SD).