How do I measure if my miRNA Mimics or Inhibitors are working?
Many researchers use a reporter system to assess the change in miRNA activity in response to the miRIDIAN Mimic or Hairpin Inhibitor treatment. The reporter construct can contain a completely complementary sequence to the mature miRNA being studied (as a control), a putative miRNA binding site, or an entire 3'UTR of a putative target gene.
In some cases, a known or putative target gene can be used for assessing miRNA activity.
miRNAs can also be detected by qRT-PCR in much the same way that mRNAs are detected. One factor to keep in mind is that you will need to add a polyA tail for the RT step if you want to use oligo dT primers. There are several commercial vendors for measuring miRNA expression by qRT-PCR so you would have to look at each particular kit for details.
For more information on this topic please see our Technical Note: Using Mimics and Inhibitors for microRNA Functional Analysis