TY - JOUR T1 - Unbiased dynamic characterization of RNA-protein interactions by OOPS JF - bioRxiv DO - 10.1101/333336 SP - 333336 AU - Rayner M. L. Queiroz AU - Tom Smith AU - Eneko Villanueva AU - Mie Monti AU - Mariavittoria Pizzinga AU - Maria Marti-Solano AU - Dan-Mircea Mirea AU - Manasa Ramakrishna AU - Robert F. Harvey AU - Veronica Dezi AU - Sven Degroeve AU - Lennart Martens AU - Gavin H. Thomas AU - Anne E. Willis AU - Kathryn S. Lilley Y1 - 2018/01/01 UR - http://biorxiv.org/content/early/2018/05/29/333336.abstract N2 - Current methods for the identification of RNA–protein interactions require a quantity and quality of sample that hinders their application, especially for dynamic biological systems or when sample material is limiting. Here, we present a new approach to enrich RNA-Binding Proteins (RBPs): Orthogonal Organic Phase Separation (OOPS), which is compatible with downstream proteomics and RNA sequencing. OOPS enables recovery of RBPs and free protein, or protein-bound RNA and free RNA, from a single sample in an unbiased manner. By applying OOPS to human cell lines, we extract the majority of known RBPs, and importantly identify additional novel RBPs, including those from previously under-represented cellular compartments. The high yield and unbiased nature of OOPS facilitates its application in both dynamic and inaccessible systems. Thus, we have identified changes in RNA-protein interactions in mammalian cells following nocodazole cell-cycle arrest, and defined the first bacterial RNA-interactome. Overall, OOPS provides an easy-to-use and flexible technique that opens new opportunities to characterize RNA-protein interactions and explore their dynamic behaviour. ER -