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

Methods. 2020 Jun 1;178:83-88. doi: 10.1016/j.ymeth.2019.09.001. Epub 2019 Sep 5.

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.

Publication types

  • Research Support, Non-U.S. Gov't