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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
Artikel, Bibliographie

Kurzbeschreibung (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.

Typ des Eintrags: Artikel
Erschienen: 2021
Autor(en): Duan, Ningjun ; Arroyo, Maria ; Deng, Wen ; Cardoso, M. Cristina ; Leonhardt, Heinrich
Art des Eintrags: Bibliographie
Titel: Visualization and characterization of RNA-protein interactions in living cells
Sprache: Englisch
Publikationsjahr: 27 Juli 2021
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Nucleic acids research
Jahrgang/Volume einer Zeitschrift: 49
(Heft-)Nummer: 18
DOI: 10.1093/nar/gkab614
Kurzbeschreibung (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.

ID-Nummer: pmid:34313753
Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
10 Fachbereich Biologie > Cell Biology and Epigenetics
Hinterlegungsdatum: 02 Aug 2021 12:28
Letzte Änderung: 25 Nov 2021 06:52
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