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In vivo detection of RNA-binding protein interactions with cognate RNA sequences by fluorescence resonance energy transfer
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SYSNO ASEP 0334099 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title In vivo detection of RNA-binding protein interactions with cognate RNA sequences by fluorescence resonance energy transfer Author(s) Huranová, Martina (UMG-J) ORCID
Jablonski, J.A. (US)
Benda, Aleš (UFCH-W) RID, ORCID
Hof, Martin (UFCH-W) RID, ORCID
Staněk, David (UMG-J) RID
Caputi, M. (US)Number of authors 6 Source Title RNA. - : Cold Spring Harbor Laboratory Press - ISSN 1355-8382
Roč. 15, č. 11 (2009), s. 2063-2071Number of pages 9 s. Language eng - English Country US - United States Keywords FRET ; FLIM ; RNA-protein interactions Subject RIV EB - Genetics ; Molecular Biology R&D Projects KAN200520801 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) LC06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) CEZ AV0Z50520514 - UMG-J (2005-2011) AV0Z40400503 - UFCH-W (2005-2011) UT WOS 000270851000014 DOI 10.1261/rna.1678209 Annotation 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. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2010
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