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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