Live-Cell RNA Imaging via Clickable TriPPPro Nucleotide Reporters.

ABSTRACT: Understanding RNA synthesis and dynamics in cells requires efficient labeling strategies that are not only compatible with cellular environments but can be performed in living cells. We developed a robust, bio-orthogonal approach for live-cell RNA labeling using TriPPPro (triphosphate prodrug) chemistry. This strategy enables the intracellular delivery of sterically demanding nucleoside triphosphates modified with inverse electron-demand Diels-Alder (IEDDA)-reactive groups, specifically trans-cyclooctene (2TCOa) and bicyclo[6.1.0]nonyne (BCN). Once hydrolyzed inside cells, these TriPPPro-modified uridines and cytidines are metabolically incorporated into nascent RNA by endogenous RNA polymerases. Subsequent IEDDA reaction with a dual-fluorogenic tetrazine-cyanine styryl dye conjugate allow

SUBMITTER: Knaack JIH 

PROVIDER: S-EPMC12790367 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature