Analysis of RBP regulation and co-regulation of mRNA 3′ UTR regions in a luciferase reporter system

Abstract

The luciferase reporter assay is a widely used tool to study the cis and trans factors controlling regulation of gene expression. In this assay, regulatory elements can be fused to the luciferase gene, and as a result effect protein output by changing rates of transcription, rates of translation, or mRNA stability. This protocol focuses on probing the function of RNA-binding proteins (RBPs) through their interactions with the 3′ untranslated region (UTR), thus examining gene expression regulation on the mRNA level. Assessment of 3′ UTR sequence requirements, as well as single and co-regulatory roles of RBPs in regulation of mRNAs will be discussed.