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Visualization and characterization of RNA-protein interactions in living cells

Duan, Ningjun ; Arroyo, Maria ; Deng, Wen ; Cardoso, M. Cristina ; Leonhardt, Heinrich (2021)
Visualization and characterization of RNA-protein interactions in living cells.
In: Nucleic acids research, 49 (18)
doi: 10.1093/nar/gkab614
Article, Bibliographie

Abstract

RNA-protein interactions are the structural and functional basis of significant numbers of RNA molecules. RNA-protein interaction assays though, still mainly depend on biochemical tests in vitro. Here, we establish a convenient and reliable RNA fluorescent three-hybrid (rF3H) method to detect/interrogate the interactions between RNAs and proteins in cells. A GFP tagged highly specific RNA trap is constructed to anchor the RNA of interest to an artificial or natural subcellular structure, and RNA-protein interactions can be detected and visualized by the enrichment of RNA binding proteins (RBPs) at these structures. Different RNA trapping systems are developed and detection of RNA-protein complexes at multiple subcellular structures are assayed. With this new toolset, interactions between proteins and mRNA or noncoding RNAs are characterized, including the interaction between a long noncoding RNA and an epigenetic modulator. Our approach provides a flexible and reliable method for the characterization of RNA-protein interactions in living cells.

Item Type: Article
Erschienen: 2021
Creators: Duan, Ningjun ; Arroyo, Maria ; Deng, Wen ; Cardoso, M. Cristina ; Leonhardt, Heinrich
Type of entry: Bibliographie
Title: Visualization and characterization of RNA-protein interactions in living cells
Language: English
Date: 27 July 2021
Journal or Publication Title: Nucleic acids research
Volume of the journal: 49
Issue Number: 18
DOI: 10.1093/nar/gkab614
Abstract:

RNA-protein interactions are the structural and functional basis of significant numbers of RNA molecules. RNA-protein interaction assays though, still mainly depend on biochemical tests in vitro. Here, we establish a convenient and reliable RNA fluorescent three-hybrid (rF3H) method to detect/interrogate the interactions between RNAs and proteins in cells. A GFP tagged highly specific RNA trap is constructed to anchor the RNA of interest to an artificial or natural subcellular structure, and RNA-protein interactions can be detected and visualized by the enrichment of RNA binding proteins (RBPs) at these structures. Different RNA trapping systems are developed and detection of RNA-protein complexes at multiple subcellular structures are assayed. With this new toolset, interactions between proteins and mRNA or noncoding RNAs are characterized, including the interaction between a long noncoding RNA and an epigenetic modulator. Our approach provides a flexible and reliable method for the characterization of RNA-protein interactions in living cells.

Identification Number: pmid:34313753
Divisions: 10 Department of Biology
10 Department of Biology > Cell Biology and Epigenetics
Date Deposited: 02 Aug 2021 12:28
Last Modified: 25 Nov 2021 06:52
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