Figure 1 : MiRNA-mediated gene-silencing assay using reporter plasmids.

From: Stability of miRNA 5′terminal and seed regions is correlated with experimentally observed miRNA-mediated silencing efficacy

Figure 1

(a) Structure of psiCHECK-SM, which has three tandem repeats in the Renilla luciferase 3′ UTR. (b) Structure of psiCHECK-CM, which has a complete-matched target sequence in the Renilla luciferase 3′ UTR. (c) The miRNA-mediated gene-silencing activities in HeLa cells as a function of miRNA concentration at 0.05, 0.5, and 5 nM. The indicated concentration of synthetic miRNA, psiCHECK-SM construct (10 ng), and pGL3-control (100 ng) were simultaneously transfected into HeLa cells, and Renilla luciferase activity / firefly luciferase activity was determined 24 hours after transfection. The gene-silencing efficiencies of target mRNA varied significantly depending on the miRNA duplex used for transfection. The sequences of miRNAs and target mRNAs are shown in Figure S1. The ordinate represents relative luciferase activity (%) and the abscissa represents miRNA concentration. The black bar indicates the result of psiCHECK-SM. The white bar, the result of psiCHECK-CM.