In vivo detection of RNA-binding protein interactions with cognate RNA sequences by fluorescence resonance energy transfer

0334099 - ÚMG 2010 RIV US eng J - Journal Article
Huranová, Martina - Jablonski, J.A. - Benda, Aleš - Hof, Martin - Staněk, David - Caputi, M.
In vivo detection of RNA-binding protein interactions with cognate RNA sequences by fluorescence resonance energy transfer.
RNA. Roč. 15, č. 11 (2009), s. 2063-2071. ISSN 1355-8382. E-ISSN 1469-9001
R&D Projects: GA AV ČR KAN200520801; GA MŠMT(CZ) LC06063
Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z40400503
Keywords : FRET * FLIM * RNA-protein interactions
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 5.198, year: 2009

Expression of the nascent RNA transcript is regulated by its interaction with a number of proteins. Misregulation of such interactions often results in impaired cellular functions that can lead to cancer and many diseases. Our understanding of RNA–protein interaction within the cellular context is thus essential for development of novel diagnostic and therapeutic tools. While there are many in vitro methods analyzing RNA–protein interactions, in vivo approaches are scarce. Here we established a method based on fluorescence resonance energy transfer (FRET), which we termed RNA-binding mediated FRET (RB-FRET). The methods enables determination of RNA–protein interaction inside cells and was tested on hnRNP H protein binding to its cognate RNA. Using two different approaches, we provided evidence that RB-FRET was sensitive enough to detect specific RNA–protein interactions in cell, providing a powerful tool to study spatial and temporal localization of specific RNA–protein complexes.
Permanent Link: http://hdl.handle.net/11104/0178922