title: A rapid method for assessing the RNA-binding potential of a protein.
abstract: In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures.
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia.
authors.forename: K
authors.initials: K
authors.lastname: Bendak
authors.forename: F E
authors.initials: FE
authors.lastname: Loughlin
authors.forename: V
authors.initials: V
authors.lastname: Cheung
authors.forename: M R
authors.initials: MR
authors.lastname: O'Connell
authors.forename: M
authors.initials: M
authors.lastname: Crossley
authors.forename: J P
authors.initials: JP
authors.lastname: Mackay
mesh_terms: D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D006493:Heparin; D015347:RNA Probes / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000032:analysis*
chemical_list: D015347:RNA Probes; D016601:RNA-Binding Proteins; D006493:Heparin