Figure 6.

Neuron-specific transduction in floxed mice by FlpO-dCre system in combination with a neurotropic PHP.eB capsid

(A) Schema showing experimental procedure. Ai75G mice, which express NLS-attached tdTomato in the presence of Cre, received intravenous injection of astrocyte-tropic AAV-F or neuron-tropic PHP.eB capsid vectors expressing Cre under control of NSE promoter, as controls. To prove the efficacy of our FlpO-dCre system, Ai75G mice similarly received a mixture of 2 NSE promoter-driven PHP.eB vectors: one expressing FlpO and another expressing dCre in the presence of FlpO. (B) Immunohistochemistry of mouse brain 3 weeks after AAV injection. Far left, fluorescence images of tdTomato (magenta) and NeuN (green) from primary motor cortex, in which square regions were enlarged and arranged at right. Arrowheads indicate tdTomato⁺ and NeuN⁻ nonneuronal cells. Scale bars in bottom left and bottom right, 100 and 10 μm, respectively. (C) Graph showing specificity (Spe.) and efficiency (Eff.) for the transduction of neurons and astrocytes. “C” and “F” below horizontal axis indicate “Cre” and “FlpO-dCre,” respectively. Data (average ± SD) were obtained from 4 mice, and the value from each mouse was plotted. n.s., not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; and ∗∗∗∗p < 0.0001 by 1-way ANOVA with Bonferroni’s post hoc test.