Validation of the miR-485-5p binding site in the BACE1 transcript. (a) The presence of the miR-485-5p target site in the 3' UTR of firefly luciferase is sufficient for depleting luciferase expression by 30%, equally effective as a perfect match positive control. The scrambled target site did not show any effect (**P-value < 0.01). This experiment was performed in HEK293T cells and each treatment consisted of 24 biological repeats; error bars represent standard error of means (SEM). (b) Over-expression of miR485-5p, but not control miRNA (miR-219) or empty vector, leads to BACE1 protein reduction by about 30% (***P-value < 0.001). Each treatment consists of 32 biological repeats and error bars represent SEM. In this experiment, the miRNA-binding site was not artificially engineered; rather, it is located in its usual place in the open reading frame of the BACE1 transcript. BACE1 protein level was measured by DiscoverRx technology.
Faghihi et al. Genome Biology 2010 11:R56 doi:10.1186/gb-2010-11-5-r56