An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNANúria Crua Asensio 1, Stefano Rizzieri 2, Alessandro Cuomo 2, Gian Gaetano Tartaglia 1,3, Elsa Zacco 1
1Center for Human Technology, Italian Institute of Technology (IIT), 2Department of Experimental Oncology, European Institute of Oncology (IEO), 3Department of Biology and Biotechnologies ‘Charles Darwin’, Sapienza University of Rome
Here, we present an optimized in vitro method to uncover, quantify, and validate protein interactors of specific RNA sequences, using total protein extract from human cells, streptavidin beads coated with biotinylated RNA, and mass spectrometry analysis.