Identification of novel dsRBPs through co-immunoprecipitation and TurboID proximity labeling

Abstract

Various non-coding RNAs, those that are not translated into a protein, are transcribed to modulate cellular pathways. dsRNA is a category of these non-coding RNAs, and those with length longer than 80bp are known to be RNAs typical of viral genome. Along with viral dsRNA which can be found in infected cells, mammalian cells also express various endogenous dsRNAs, most of which still has unknown functional roles. In this research, we have used co-immunoprecipitation (co-IP) and TurboID proximity labeling (PL) to extensively find those proteins that can interact with the endogenous dsRNAs. Co-IP is a method whereby a specific antibody is used to separate a specific target protein complex. PL technique utilizes promiscuous enzyme to label all proteins within a specific radius proximal to the enzyme. By comparing the proteomic profile from the two experiments, we aim to newly identify dsRBPs that were previously unrecognized, and further study its role in human cellular pathways.