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

Modulation of human miR-1225 abundance using steric-blocking ASOs. (A) ASO sequences and target site on PKD1 RNA are shown. (B) Radiolabeled stem–loop RT-PCR (top, miR-1225/miR-16) and RT-PCR (bottom, PKD1/GAPDH) of RNA isolated from HeLa cells that were transiently transfected with a WT human PKD1 minigene and 50 nM ASO, or minigene alone (mock) as a control. GAPDH and miR-16 are controls for normalization of total RNA quantity among samples. Intron 45 retained indicates amplicon from RNA with unspliced intron 45. (C) Quantitation of spliced PKD1 mRNA and miR-1225 normalized to GAPDH and miR-16 values, respectively, and graphed relative to mock-treated sample.