RNA‐Targeting Carbon Dots for Live‐Cell Imaging of Granule Dynamics

Abstract It is significant to monitor the different RNA granules dynamics and phase separation process inside cells under various stresses, for example, oxidative stress. The current small‐molecule RNA probes work well only in fixed cells and usually encounter problems such as insufficient stability and biocompatibility, and thus a specific RNA‐targeting fluorescent nanoprobe that can be used in the living cells is urgently desired. Here, the de novo design and microwave‐assisted synthesis of a novel RNA‐targeting, red‐emissive carbon dots (named as M‐CDs) are reported by choosing neutral red and levofloxacin as precursors. The as‐synthesized M‐CDs is water‐soluble with a high fluorescence quantum yield of 22.83% and can selectively bind to RNA resulting in an enhanced red fluorescence. More interestingly, such an RNA‐targeting, red‐emissive M‐CDs can be fast internalized into cells within 5 s and thus used for real‐time imaging the dynamic process of intracellular stress granules under oxidative stress, revealing some characteristics of granules that have not been identified by previously reported RNA and protein biomarkers. This research paves a new pathway for visualizing bulk RNA dynamics and studying phase‐separation behaviors in living cells by rational design of the fluorescent carbon dots in terms of structure and functionality.