Manipulating MiRNA Expression to Uncover Hidden Functions.
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.
Department of Tumor Biology, H. Lee Moffitt Cancer Center and Research Institute, SRB-3, 12902 Magnolia Drive, Tampa, FL, 33612, USA. Sinead.Aherne@moffitt.org.|National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland. Sinead.Aherne@moffitt.org.
Sinéad T
ST
Aherne
National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland.
Nga T
NT
Lao
D001483:Base Sequence; D002460:Cell Line; D055785:Gene Knockdown Techniques; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D015533:Transcriptional Activation; D014162:Transfection
D035683:MicroRNAs
Cloning; Ligation; Transfection; miRNA-Target