Intramolecular Proximity-Induced Amplification for Accurate Imaging of Glycosylated RNAs in Living Cells and Zebrafish

Abstract

As emerging molecules, glycosylated RNAs (glycoRNAs) have been demonstrated to participate in cellular communication, developmental signaling pathway regulation, and immune recognition. Revealing the dynamics of glycoRNAs during biological development is crucial for advancing our understanding of these complex processes. However, to date, the dynamic changes in glycoRNAs during biological development have not been investigated due to the lack of visualization methods with high contrast and high sensitivity for monitoring glycoRNAs in complex tissues. Herein, we developed an intramolecular proximity-induced amplification (IPIA) strategy based on the dual-specific recognition sites of RNA molecules and sialic acid for accurate visualization of glycoRNAs in situ. Employing its multisite cooperative recognition capacity and excellent capture performance, the IPIA strategy successfully mapped the spatiotemporal dynamics of glycoRNAs during zebrafish development, while also setting the stage for elucidating their roles in organogenesis. The IPIA strategy provides a new tool for unveiling the roles of glycoRNAs in critical biological events and exploring their potential applications in disease diagnosis and therapy.