Title: "Illuminating RNA Dynamics with Riboglow, One Molecule at a Time"
Esther Braselmann, Research Associate, Biofrontiers Institute, University of Colorado Boulder
Spatiotemporal dynamics of coding and non-coding RNAs play central roles in biology, and visualizing these processes on a single cell level may enable novel insights in underlying biology. RNA visualization has lagged behind the protein field, and there is a need to develop tools to label and visualize RNA of interest in live cells over time. We developed ‘Riboglow’ to visualize RNA live in mammalian cells that combines small size, robust applicability and multi-color fluorescent tags. A small RNA tag is attached as a fusion to an RNA of interest and addition of an organic probe induces fluorescence. We have validated Riboglow as a robust platform to visualize and track RNA dynamics in diverse applications in live mammalian cells. First, we visualized live mRNA recruitment to stress granules. Second, we tagged the non-coding U1 snRNA with Riboglow, and demonstrated its recruitment to cytosolic granules, confirming versatility to tag and visualize small non-coding RNAs that are not compatible with existing RNA imaging systems. Third, we confirmed that single mRNAs can be detected and tracked with Riboglow, and their dynamics can be quantified. Most recently, we multiplexed tracking of Riboglow-tagged mRNA with orthogonal fluorescence labeling systems to quantify mRNA dynamics during protein translation. Together, we developed a small and versatile platform to tag and visualize RNA that is compatible with diverse RNAs of interest for live cell imaging while only minimally disrupting function.