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

Modulating mouse miR-1225 abundance with ASOs. (A) ASO sequences and target sequences are shown. (B) Mouse 89N cells were transiently transfected with 50 nM of ASO or lipofectamine alone (mock) as a control. miR-1225 abundance and Pkd1 splicing were assessed by radiolabeled stem–loop RT-PCR and RT-PCR, respectively. Gapdh, snoRNA65, and miR-877 are controls for loading. C indicates a mock transfection where no ASO was added, and – indicates untransfected cells. (C) Graph represents quantitation of amplicons in (B). miR-1225 abundance was normalized miR-877 and graphed relative to mock-treated cells. Pkd1 abundance is normalized Gapdh and graphed relative to mock-treated sample.