Elevating microRNA levels by targeting biogenesis with steric-blocking antisense oligonucleotides

ASO-16 restores miR-1225 derived from the ADPKD variant IVS45-14T > C. (A) Stem–loop RT-PCR and RT-PCR analysis of miR-1225 and PKD1 mRNA, respectively, of RNA isolated from HEK-293T cells transiently transfected with plasmids expressing WT, IVS45-14T > C, IVS45 + 34_27del14, IVS45-36_-12del25, or IVS45-3C > G PKD1 minigenes with (+) or without (−) ASO-16 cotransfection. (B) Quantitation of miR-1225 and PKD1 normalized to miR-16 and GAPDH, respectively. Error bars represent SEM, n = 4, (*) P<0.05 multiple paired t-test comparing control (C) and ASO-16 for each minigene.