Imaging RNA and RNA biology using RNA mimics of green fluorescent protein (347.1)

Abstract

Green fluorescent protein (GFP) and its derivatives have transformed the use and analysis of proteins for diverse applications. Like proteins, RNA has complex roles in cellular function and is increasingly used for various in vitro and in vivo applications, but a comparably robust and simple approach for fluorescently tagging RNA is lacking. We will describe the generation of RNA aptamers that bind fluorophores resembling the fluorophore in GFP. These RNAs activate the fluorescence of these fluorophores, resulting in a palette of RNA‐fluorophore complexes that span the visible spectrum. An RNA‐fluorophore complex resembling enhanced GFP (EGFP), termed Spinach, emits a green fluorescence comparable in brightness to fluorescent proteins. Spinach is markedly resistant to photobleaching, and Spinach fusion RNAs can be used to image RNA localization and other RNA regulatory processes in cells. We will also discuss the use of Spinach to image other biological molecules in cells. These RNA mimics of GFP provide novel approaches to image RNA biology and other processes in cells.