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Related Experiment Video

Updated: Jan 6, 2026

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
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Live-Cell RNA Imaging via Clickable TriPPPro Nucleotide Reporters.

J Iven H Knaack1, Eileen List2, Dörte Stalling3,4,5,6

  • 1Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146, Hamburg, Germany.

Angewandte Chemie (International Ed. in English)
|November 28, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed TriPPPro (triphosphate prodrug) chemistry for efficient live-cell RNA labeling. This bio-orthogonal method allows real-time imaging of newly synthesized RNA with high specificity and cellular compatibility.

Keywords:
Click‐chemistryLive‐cell ImagingNucleotidesRNA LabelingTriPPPro‐compounds

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Efficient labeling of RNA in living cells is crucial for understanding cellular processes.
  • Existing methods often face limitations in cellular compatibility and labeling efficiency.

Purpose of the Study:

  • To develop a robust, bio-orthogonal approach for live-cell RNA labeling.
  • To enable real-time imaging of nascent RNA synthesis using a novel chemical strategy.

Main Methods:

  • Development of TriPPPro (triphosphate prodrug) chemistry for intracellular delivery of modified nucleoside triphosphates.
  • Utilizing inverse electron-demand Diels-Alder (IEDDA) click chemistry for subsequent RNA labeling.
  • Employing dual-fluorogenic tetrazine-cyanine styryl dye conjugates for high-contrast imaging.

Main Results:

  • TriPPPro successfully delivered sterically demanding nucleoside triphosphates into cells.
  • Metabolic incorporation of modified uridines and cytidines into nascent RNA by endogenous RNA polymerases.
  • Achieved wash-free, high-contrast imaging of RNA synthesis with nucleolar localization and specificity for newly transcribed RNA.
  • Demonstrated superior labeling efficiency and cellular compatibility compared to conventional transporter-based systems.

Conclusions:

  • TriPPPro chemistry provides a modular, non-genetic, and highly specific method for real-time RNA imaging.
  • This platform has broad applicability in RNA biology research and antiviral studies.
  • The developed strategy overcomes limitations of existing live-cell RNA labeling techniques.