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Fluorogenic Covalent Probes for RNA via 2′-Hydroxyl Acylation with High Selectivity
Dec 25, 2025

Stanford Team Unveils New Fluorogenic Probes for Selective RNA Imaging

A research team led by Prof. Eric T. Kool at Stanford University has developed a novel class of covalent fluorogenic probes—dubbed RiboLight (RL) fluorophores—that selectively label RNA through reaction with its 2′-hydroxyl (2′‑OH) groups. Published in the Journal of the American Chemical Society, the work presents a sequence-independent, wavelength-tunable strategy that delivers up to 390-fold fluorescence enhancement and 970-fold selectivity for RNA over DNA.

Unlike conventional non-covalent dyes, RL probes operate under mild aqueous conditions, minimize background noise, and enable clear RNA visualization in gels, solution, and living cells. Crucially, the labeling is reversible under gentle conditions, allowing recovery of native RNA for downstream analysis.By exploiting RNA’s unique 2′‑OH group—a feature absent in DNA—the technology overcomes longstanding challenges in RNA-specific imaging and opens new avenues for real-time tracking and molecular analysis of RNA in complex biological systems.

Development of RNA-selective fluorogenic covalent acylating probes (RiboLight-RL)


 


References

[1] Shin J, Kim M J, Kool E T. Fluorogenic Covalent Probes for RNA[J]. J. Am. Chem. Soc. 2025. https://doi.org/10.1021/jacs.5c14938
 

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