MicroRNA (miRNA) functional analyses can be performed by using miRNA inhibitors (Anti-miR™ miRNA Inhibitors) and synthetic miRNA mimics (Pre-miR™ siRNA Precursors), which are now available for most known miRNAs. Anti-miR and Pre-miR positive, negative, and labeled controls have been designed to serve as convenient and effective controls for experiments involving Anti-miR miRNA Inhibitors and Pre-miR miRNA Precursors.
Positive Control: Pre-miR™ hsa-miR-1 miRNA Precursor
Pre-miR hsa-miR-1 miRNA Precursor, which mimics mature miR-1, has been shown to down-regulate the Protein Tyrosine Kinase 9 (PTK9; twinfillin) gene at the mRNA level . Expression of PTK9 mRNA is decreased by 60–95% following Pre-miR hsa-miR-1 miRNA Precursor transfection, compared to transfection with Pre-miR miRNA Precursor–Negative Control #1, using real-time PCR and TaqMan® Gene Expression Assays [Assay ID#s Hs00702289_s1 (human) and Mm01598980_g1 (mouse); data not shown]. Therefore, Pre-miR hsa-miR-1 miRNA provides a convenient positive control for miRNA-mediated gene silencing of PTK9, when monitored using the appropriate TaqMan Gene Expression Assays and real-time PCR. The Pre-miR positive control can be used to confirm that the transfection procedure in cell cultures support miRNA-mediated gene silencing.
Anti-miR™ and Pre-miR™ miRNA Negative Controls
Anti-miR miRNA Inhibitor–Negative Control #1 is a nontargeting sequence Anti-miR molecule designed to serve as a negative control for experiments involving Anti-miR miRNA Inhibitors. These Anti-miR miRNA negative controls are transfected using the same methods as those for experimental Anti-miR miRNA Inhibitors.
Pre-miR miRNA Precursor–Negative Control #1 and #2 are double-stranded RNA oligonucleotides with nontargeting sequences, which are designed to serve as negative controls for experiments involving Pre-miR miRNA Precursors.