Lost In Translation: Investigating Translational Repression With Optogenetic RNA Clustering

Abstract

RNA translation is a highly regulated and integral process for all living cells. In recent years, researchers have been investigating the role of phase separated membraneless organelles – where RNA and proteins can co-localized – in translational control. These RNA-protein bodies (RNPs) in and of themselves are a diverse system that plays roles in single cells as well as in developing organisms. While many studies have been conducted on RNPs in vitro, little research has been performed to investigate their linkage with translational regulation in live cells. Here we applied optogenetics to create a controllable RNA-protein clustering system in NIH 3T3 cells to see whether RNP phase separation can regulate translation. We then quantified the translation of a fluorescent reporter in the presence and absence of clustering. The building of this controllable RNA-protein clustering system allows for further investigations into the role of phase separation in RNA translational regulation and development.