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A versatile tandem RNA isolation procedure to capture in vivo formed mRNA-protein complexes

Matia-González, AM, Iadevaia, Valentina and Gerber, Andre (2017) A versatile tandem RNA isolation procedure to capture in vivo formed mRNA-protein complexes Methods, 118-9. pp. 93-100.

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Abstract

We describe a tandem RNA isolation procedure (TRIP) that enables purification of in vivo formed messenger ribonucleoprotein (mRNP) complexes. The procedure relies on the purification of polyadenylated mRNAs with oligo(dT) beads from cellular extracts, followed by the capture of specific mRNAs with 3'-biotinylated 2'-O-methylated antisense RNA oligonucleotides, which are recovered with streptavidin beads. TRIP was applied to isolate in vivo crosslinked mRNP complexes from yeast, nematodes and human cells for subsequent analysis of RNAs and bound proteins. The method provides a basis for adaptation to other types of polyadenylated RNAs, enabling the comprehensive identification of bound proteins/RNAs, and the investigation of dynamic rearrangement of mRNPs imposed by cellular or environmental cues.

Item Type: Article
Subjects : Biosciences and Medicine
Divisions : Faculty of Health and Medical Sciences > School of Biosciences and Medicine
Authors :
NameEmailORCID
Matia-González, AMUNSPECIFIEDUNSPECIFIED
Iadevaia, Valentinav.iadevaia@surrey.ac.ukUNSPECIFIED
Gerber, Andrea.gerber@surrey.ac.uk0000-0002-2398-6592
Date : 15 April 2017
Identification Number : 10.1016/j.ymeth.2016.10.005
Copyright Disclaimer : © 2016. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
Uncontrolled Keywords : RNA-binding protein; RNA-protein interaction; ribonucleoprotein complex; posttranscriptional regulation; antisense oligonucleotides; non-coding RNA
Depositing User : Symplectic Elements
Date Deposited : 21 Oct 2016 13:25
Last Modified : 07 Jul 2017 14:23
URI: http://epubs.surrey.ac.uk/id/eprint/812544

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