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Transcript specific mRNP capture from Drosophila egg-chambers for proteomic analysis.

Authors
  • Wippich, Frank1
  • Ephrussi, Anne2
  • 1 Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany. Electronic address: [email protected] , (Germany)
  • 2 Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany. , (Germany)
Type
Published Article
Journal
Methods
Publisher
Elsevier
Publication Date
Jun 01, 2020
Volume
178
Pages
83–88
Identifiers
DOI: 10.1016/j.ymeth.2019.09.001
PMID: 31493515
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

mRNA binding proteins (RBPs) play a major role in post-transcriptional control of gene expression. To understand the complex regulatory processes regulating a specific mRNA during its life-time, a comprehensive view of the bound RBPs is essential. Here, we describe a method for transcript-specific isolation of endogenous ribonucleoprotein complexes (RNPs) from Drosophila egg-chambers. The method, which is based on in-solution hybridization of short biotinylated antisense DNA oligonucleotide probes to multiple segments of a transcript of interest allows unbiased identification of associated proteins by quantitative proteomics. Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.

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