CBRPP: a new RNA-centric method to study RNA-protein interactions

• Published in: RNA biology Vol. 18; no. 11; pp. 1608 - 1621
• Main Authors: Li, Yunfei, Liu, Shengde, Cao, Lili, Luo, Yujie, Du, Hongqiang, Li, Siji, Zhang, Zeming, Guo, Xuefei, Tian, Wenmin, Wong, Catherine Cl, You, Fuping
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 02.11.2021
• Subjects: Biotinylation; HEK293 Cells; Humans; Mass Spectrometry - methods; Protein Binding; Protein Interaction Mapping; RNA - genetics; RNA - metabolism; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Staining and Labeling; Technical Paper; dPspCas13b; RNA-protein interactions; turboid; bioid2; apex2; dRfxCas13d; basu
• ISSN: 1547-6286; 1555-8584
• DOI: 10.1080/15476286.2021.1873620

RNA and protein are interconnected biomolecules that can influence each other's life cycles and functions through physical interactions. Abnormal RNA-protein interactions lead to cell dysfunctions and human diseases. Therefore, mapping networks of RNA-protein interactions is crucial for understanding cellular processes and pathogenesis of related diseases. Different practical protein-centric methods for studying RNA-protein interactions have been reported, but few robust RNA-centric methods exist. Here, we developed CRISPR-based RNA proximity proteomics (CBRPP), a new RNA-centric method to identify proteins associated with an endogenous RNA of interest in native cellular context without pre-editing of the target RNA, cross-linking or RNA-protein complexes manipulation . CBRPP is based on a fusion of dCas13 and proximity-based labelling (PBL) enzyme. dCas13 can deliver PBL enzyme to the target RNA with high specificity, while PBL enzyme labels the surrounding proteins of the target RNA, which are then identified by mass spectrometry.