Evolving Methods in Defining the Role of RNA in RNP Assembly

Abstract

The causative connection between aberrant liquid–liquid phase separation (LLPS) of ribonucleoprotein (RNP) granules and neurodegeneration is well known. LLPS driven by intrinsically disordered proteins has been intensely investigated. However, the role of RNA and RNA–protein interaction in RNP granule assembly, properties, maintenance, and eventual onset and progression of neurodegeneration remains poorly understood. A critical hurdle in addressing this question is the shortcomings of currently employed ensemble methods in probing very early-stage molecular events. Here, we present a unique combination of single-molecule biophysical and ensemble methods that can dissect single protein–RNA interaction. The advantage of this approach is that it provides a potential tool to identify early-stage molecular defects that may contribute to the onset of neurodegenerative diseases.