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Verfasst von:Rogell, Birgit [VerfasserIn]   i
 Fischer, Bernd [VerfasserIn]   i
 Krijgsveld, Jeroen [VerfasserIn]   i
 Castello, Alfredo [VerfasserIn]   i
 Hentze, Matthias W. [VerfasserIn]   i
Titel:Specific RNP capture with antisense LNA/DNA mixmers
Verf.angabe:Birgit Rogell, Bernd Fischer, Mandy Rettel, Jeroen Krijgsveld, Alfredo Castello, Matthias W. Hentze
Jahr:2017
Umfang:13 S.
Fussnoten:Gesehen am 15.10.2018 ; Published in Advance May 5, 2017
Titel Quelle:Enthalten in: RNA
Ort Quelle:Cold Spring Harbor, NY : Laboratory Press, 1995
Jahr Quelle:2017
Band/Heft Quelle:23(2017), 8, Seite 1290-1302
ISSN Quelle:1469-9001
Abstract:RNA-binding proteins (RBPs) play essential roles in RNA biology, responding to cellular and environmental stimuli to regulate gene expression. Important advances have helped to determine the (near) complete repertoires of cellular RBPs. However, identification of RBPs associated with specific transcripts remains a challenge. Here, we describe “specific ribonucleoprotein (RNP) capture,” a versatile method for the determination of the proteins bound to specific transcripts in vitro and in cellular systems. Specific RNP capture uses UV irradiation to covalently stabilize protein-RNA interactions taking place at “zero distance.” Proteins bound to the target RNA are captured by hybridization with antisense locked nucleic acid (LNA)/DNA oligonucleotides covalently coupled to a magnetic resin. After stringent washing, interacting proteins are identified by quantitative mass spectrometry. Applied to in vitro extracts, specific RNP capture identifies the RBPs bound to a reporter mRNA containing the Sex-lethal (Sxl) binding motifs, revealing that the Sxl homolog sister of Sex lethal (Ssx) displays similar binding preferences. This method also revealed the repertoire of RBPs binding to 18S or 28S rRNAs in HeLa cells, including previously unknown rRNA-binding proteins.
DOI:doi:10.1261/rna.060798.117
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext: http://dx.doi.org/10.1261/rna.060798.117
 Volltext: http://rnajournal.cshlp.org/content/23/8/1290
 DOI: https://doi.org/10.1261/rna.060798.117
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:ribosomal RNA
 RNA interactome
 RNA-binding proteins
 RNA-protein interactions
 specific RNP capture
K10plus-PPN:1581914547
Verknüpfungen:→ Zeitschrift

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