RNA-scaffolded RNP condensation, its physiologic regulation, and therapeutic modulation

Ribonucleoprotein complexes (RNPs) play key roles in RNA regulation. Many mRNAs, especially those localised to neuronal axons or dendrites, promote the condensation of RNPs into microscopically visible complexes, often referred to as ‘RNA granules’. Mutations in RNA-binding proteins (RBPs) that cause diseases such as amyotrophic lateral sclerosis (ALS) often affect the biophysical properties of RNP condensation. Most work to date has focused on protein-protein interactions within condensates. Recent discoveries however, indicate that long RNA molecules can provide scaffolds for RNP condensation via their secondary structures, methylation status and repetitive, cooperative interactions with collections of RBPs. Here, we will build on our collaborative experimental and computational experience in high-throughput RNA technologies to uncover the principles underlying RNP condensation, with a focus on the role of long-range RNA secondary structures in vivo. We will establish new approaches based on CRISPR-Cas13 to modulate RNP condensation on NEAT1, a well-known RNA scaffold. We shall then apply these techniques together with computational modelling to interrogate the roles of long-range RNA structures in RNP condensation on 3’UTRs, and thus in the regulation of localised mRNA activities. Finally, we will assess whether, by modulating the structured RNA scaffolds, we can neutralise aberrant RNP condensation in ALS.