In Situ Assembly of Fluorogenic RNA for Screening Natural Anti‐Liver Fibrosis Products via Dynamic Visualization of COL1A1 mRNA

Abstract Liver fibrosis, a critical precursor to cirrhosis and a leading cause of mortality, highlights the urgent need for the identification of effective therapeutics. Activation of hepatic stellate cells (HSCs) is a key process in liver fibrosis. This study presents a live‐cell drug‐screening approach that specifically targets fibrosis‐associated collagen type I alpha 1 ( COL1A1 ) mRNA in activated HSCs through the use of fluorogenic RNA self‐assembly. It employs a dual‐probe system to construct an RNA Mango II structure, which upon binding with COL1A1 mRNA, facilitates activation of the TO1‐Biotin fluorophore, thereby enabling the visualization of mRNA to indicate HSC activation levels. Through the application of a high throughput live‐cell screening system, dihydrotanshinone I (DHT) is identified as potent leading antifibrotic compound, evidenced by its inhibitory effects on COL1A1 mRNA expression. The therapeutic efficacy of DHT is further substantiated by monitoring COL1A1 mRNA dynamics following treatment. In vivo studies demonstrates the sustained administration of DHT significantly ameliorated liver fibrosis in mice models. This method offers a simple, cost‐effective approach of visualizing RNA dynamics and conducting drug screening in live cells, presenting a significant potential for the development of hepatic fibrosis therapies.