서강대학교

초록 (요약문)

RNA is commonly known as the molecule that carries genetic information from DNA to protein. However, recently RNA has been receiving a-great attention for other roles. It can act as a catalyst for various metabolic processes and store genetic information itself. As a result, a lot of research is going on to utilize RNA in various metabolic pathways to develop RNA-based therapeutics. In this study, we aimed to develop a technique to visualize synthesized mRNA in artificial cells without additional treatments such as cell lysis or washing. To accomplish this, we employed 5-Ethynyl Uridine (EU) as an RNA labeling nucleoside analog and used click chemistry to attach a fluorescent material, 3-azido-7-hydroxycoumarin for microscopic visualization. Coumarin only has fluorescence when forming triazole rings through copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) reaction. This allows for selective visualization of mRNA in artificial cells. We designed an experiment to develop the click-chemistry protocol that could selectively induce the reaction between EU and coumarin in artificial cell conditions. Additionally, we applied this protocol to visualize mRNA and successfully detected mRNA in GUV using confocal microscopy. This click-chemistry-based visualization method provides a novel approach to selectively detect biomolecules within liposomes, including artificial cells, with high selectivity and strong fluorescence intensity.