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ReTagX: Un sistema de etiquetas fluorescentes regenerativas para nanoscopía y seguimiento de moléculas únicas en

Dongjuan Si¹, Zhihao Dong¹, Gang Xing¹

¹MOE Key Laboratory of Smart Drug Delivery, MOE Innovative Center for New Drug Development of Immune Inflammatory Diseases, School of Pharmacy, Endoscopy Center and Endoscopy Research Institute, Zhongshan Hospital, Fudan University, Shanghai, China.

Nature communications

|December 12, 2025

Abstract:

Live-cell fluorescence nanoscopy and single-molecule tracking offer profound insights into cellular biology, but their application is limited by fluorescent tags with low brightness, large size, and irreversible photobleaching. Here, we present ReTagX, a bright, regenerative, compact array tag system enabling rapid recovery of photobleached signals through reversible, fluorogenic labeling. Newly developed ReTag1 (~18 kDa) with highly fluorogenic MaP655-TMP achieves a signal-to-noise ratio up to 204-fold and allows fluorescence recovery within 1 minute in wash-free live-cell imaging. This tag extends STED imaging of mitochondrial TOM20 from fewer than 5 to over 300 frames, enabling long-term visualization of mitochondrial dynamics at 80 nm resolution for over 100 minutes. Multimeric ReTag1 arrays, such as ReTag8 (~156 kDa), increase brightness 5.5-fold and extend TNFR2 single-molecule tracking ~2.9-fold, preserving >80% of trajectories over 5 minutes. ReTagX provides a robust platform for high-resolution, long-term molecular visualization of cellular dynamics in living cells.

Palabras clave:

etiquetas fluorescentes nanoscopía seguimiento de moléculas únicas células vivas regeneración fotoblanqueo imagen de alta resolución

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