As a further test of the correlation between miR-34a levels and its putative endogenous target genes in human iPSC-derived neurons, we used a miRNA inhibitor construct (anti-miR-34a) designed through its sequence complementarity to selectively trap endogenous miR-34a thereby preventing miR-34a association with its target genes along with a non-targeted control inhibitor construct (anti-scrambled). As a result of inhibiting endogenous miR-34a, expression of a miR-34a inhibitor construct is expected to reduce effects of miR-34a on mRNA stability and translational suppression resulting in an increase in target gene expression. The anti-miR-34a and anti-scrambled constructs were expressed in hNPCs from both the healthy control and BD son patient. After 2 weeks of differentiation, as shown in Figure 2F, as predicted, expression of the anti-miR34a construct increased the expression of both ANK3 and CACNB3 in both the control and BD patient iPSC-derived neurons.
