Simultaneous observation of RNAs and their binding proteins in plant cells

Abstract

RNA is key component of a versatile group of complexes that play diverse roles in a wide range of biological processes, including plant acclimation to ever-changing environments. From synthesis to degradation, RNA molecules interact with cognate proteins that assist in processes such as transcription, splicing, modification, trafficking, and the execution of their functions. While numerous valuable techniques exist to study RNA-protein interactions, visualizing RNAs and their associated proteins simultaneously within cells remains challenging, despite its potential to provide deeper insights into the biology of RNA-protein interactions. In this study, we adapted a modified immunofluorescence (IF) assay in combination with RNA fluorescence in situ hybridization (FISH) to successfully visualize the colocalization of endogenous as well as infectious RNAs with their cognate binding proteins in plant nucleus. This new method that combines IF and FISH will facilitate future studies on RNA and protein colocalization in various plant systems.