A Live-Cell NanoBRET Assay to Monitor RNA-Protein Interactions and Their Inhibition by Small Molecules.

ABSTRACT: RNA-protein interactions are critical for cellular processes, including translation, pre-mRNA splicing, post-transcriptional modifications, and RNA stability. Their dysregulation is implicated in diseases such as myotonic dystrophy type 1 (DM1) and amyotrophic lateral sclerosis (ALS). To investigate RNA-protein interactions, here is described a live-cell NanoBioluminescence Resonance Energy Transfer (NanoBRET) assay to study the interaction between expanded r-(CUG) repeats [r-(CUG)^exp] and muscleblind-like 1 (MBNL1), central to DM1 pathogenesis. This r-(CUG)^exp sequesters MBNL1, a regulator of alternative pre-mRNA splicing, in nuclear foci causing splicing dysregulation. In the NanoBRET assay, r-(CUG)^exp acts as a scaffold to bring into proximity a BRET pai

SUBMITTER: Shan J 

PROVIDER: S-EPMC12670313 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature