There are a numerous target prediction algorithms that allow users to identify putative targets of their microRNAs (miRNAs) of interest. Although these tools are useful to gain insight into the potential role of miRNAs in regulating cellular processes, physical manipulation of the expression of the miRNA is the most superior way to truly determine the function of the miRNA in the system of interest. This chapter outlines methods to reveal miRNA function by modulating miRNA expression by transient transfection of miRNA mimics and inhibitors, and stable overexpression and reduction of miRNA expression using plasmid overexpression and sponge vectors.
Keywords: Cloning; Ligation; Transfection; miRNA-Target.