Enzymatic RNA Biotinylation for Affinity Purification and Identification of RNA-Protein Interactions.

ABSTRACT: Throughout their cellular lifetime, RNA transcripts are bound to proteins, playing crucial roles in RNA metabolism, trafficking, and function. Despite the importance of these interactions, identifying the proteins that interact with an RNA of interest in mammalian cells represents a major challenge in RNA biology. Leveraging the ability to site-specifically and covalently label an RNA of interest using E. coli tRNA guanine transglycosylase and an unnatural nucleobase substrate, we establish the identification of RNA-protein interactions and the selective enrichment of cellular RNA in mammalian systems. We demonstrate the utility of this approach through the identification of known binding partners of 7SK snRNA via mass spectrometry. Through a minimal 4-nucleotide mutation of the lon

SUBMITTER: Busby KN 

PROVIDER: S-EPMC10198264 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature