Lymphocytic Leukemias

pH1ant-miR-20a[Artifically engineered RNA]

miR-20a[miRNA]

E2F1[mRNA]

K562 Cells

Mus musculus (mouse)

Nucleic Acids Res 2007 35, e149 doi:10.1093/nar/gkm971 PMID:18025036

Lentivirus-mediated antagomir expression for specific inhibition of miRNA function

Western blot

Here, we describe the induction of stable loss-of-function phenotypes for specific miRNAs by lentivirus-mediated antagomirexpression. Lentivirally expressed antagomirs are transcribed from a H1-promoter located within the lentiviral 3LTR and were directed against miRNAs encoded on the polycistronic miR17-92 transcript. Functional silencing of miR-18a, miR-19b and miR-20a by the corresponding antagomirs specifically relieves miRNA-mediated reporter gene repression. Inhibition of miRNA function correlates to reduction of miRNA amplification by miRNA-specific quantitative RT-PCR. Furthermore, protein expression of E2F1, a known miR-20 target, is enhanced by lentivirally expressed anti-miR-20 antagomirs in a dose-dependent manner, whereas over-expression of miR-20a reduces E2F1 levels.