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Methods. 2019 Sep 5. pii: S1046-2023(18)30489-4. doi: 10.1016/j.ymeth.2019.09.001. [Epub ahead of print]

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

Author information

1
Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany. Electronic address: frank.wippich@embl.de.
2
Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

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.

KEYWORDS:

Maternally deposited mRNAs; Post-transcriptional regulation; RNA binding protein; RNA-protein interaction; mRNP capture

PMID:
31493515
DOI:
10.1016/j.ymeth.2019.09.001
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