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Updated: Feb 9, 2026

An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA
Published on: February 17, 2023
RBPscan: A quantitative in vivo tool for profiling RNA-binding protein interactions.
Dmitry A Kretov1, Owen Sanborn2, Thora McIsaac2
1Department of Biochemistry and Cell Biology, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USA; Department of Molecular Biology, Medical Biochemistry, and Pathology, Faculty of Medicine, Université Laval, Québec, QC G1V 0A6, Canada; Oncology Division, CHU de Québec - Université Laval Research Center, Québec, QC G1J 0J9, Canada; Université Laval Cancer Research Center, Québec, QC G1J 0J9, Canada.
A new method, RBPscan, uses RNA editing to profile RNA-binding protein (RBP) interactions in vivo. This tool quantifies RBP binding strength and identifies binding motifs across species.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- RNA-binding proteins (RBPs) regulate gene expression post-transcriptionally.
- Quantitative analysis of RBP-RNA interactions in vivo is challenging.
Purpose of the Study:
- To develop a novel method for profiling RBP-RNA interactions in vivo.
- To quantify RBP binding strength, dissociation constants, and binding motifs.
Main Methods:
- Developed RBP specificity and contextual analysis via nucleotide editing (RBPscan).
- Integrated RNA editing with massively parallel reporter assays.
- Fused RBPs to the adenosine deaminase acting on RNA (ADAR) catalytic domain.
Main Results:
- RBPscan successfully quantified RBP binding strength and dissociation constants.
- Identified RBP binding motifs and mapped Pumilio-binding sites in NORAD.
- Demonstrated utility in zebrafish, human cells, and yeast.
Conclusions:
- RBPscan is a versatile tool for studying RBP-RNA interactions.
- Complements existing methods for post-transcriptional regulatory network analysis.
- Enables quantitative insights into RBP function across diverse systems.
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