Adaptation of RiPCA for the Live-Cell Detection of mRNA-Protein Interactions

Abstract

ABSTRACTRNA-binding proteins (RBPs) act as essential regulators of cell fate decisions through their ability to bind and regulate the activity of cellular RNAs. For protein-coding messenger RNAs (mRNAs), RBPs control the localization, stability, degradation, and ultimately translation of mRNAs to impact gene expression. Disruption of the vast network of mRNA-protein interactions has been implicated in many human diseases, and accordingly, targeting these interactions has surfaced as a new frontier in RNA-targeted drug discovery. To catalyze this new field, methods are needed to enable the detection and subsequent screening of mRNA-RBP interactions, particularly in live cells. Using our laboratory’s RNA-interaction with Protein-mediated Complementation Assay (RiPCA) technology, herein we describe its application to mRNA-protein interactions and present a guide for the development of future RiPCA assays for structurally diverse classes of mRNA-protein interactions.