title: RNA Whole-Mount In Situ Hybridization Proximity Ligation Assay (rISH-PLA), an Assay for Detecting RNA-Protein Complexes in Intact Cells.
abstract: Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein interactions as a consequence of the complexities associated with working with RNA. This unit describes a method for the adaptation of the In Situ Hybridization-Proximity Ligation Assay (ISH-PLA) to the study of RNA regulation (rISH-PLA). The rISH-PLA assay allows the identification of a given RNA-protein complex at subcellular and single-cell resolution, thus avoiding the lack of spatial resolution and sensitivity associated with assaying heterogeneous cell populations from which conventional RNA-protein interaction detection techniques suffer. This technique will be particularly usefully for studying the activity of RNA binding proteins (RBPs) in complex mixtures of cells, for example tissue sections or whole embryos. © 2017 by John Wiley & Sons, Inc.
authors.affiliation: Biophysics Laboratories, Institute of Biomedical and Biomolecular Sciences, University of Portsmouth, Portsmouth, United Kingdom.
authors.forename: Ioannis M
authors.initials: IM
authors.lastname: Roussis
authors.affiliation: Biophysics Laboratories, Institute of Biomedical and Biomolecular Sciences, University of Portsmouth, Portsmouth, United Kingdom.
authors.forename: Fiona A
authors.initials: FA
authors.lastname: Myers
authors.affiliation: Biophysics Laboratories, Institute of Biomedical and Biomolecular Sciences, University of Portsmouth, Portsmouth, United Kingdom.
authors.forename: Garry P
authors.initials: GP
authors.lastname: Scarlett
mesh_terms: D000818:Animals; D017403:In Situ Hybridization / Q000379:methods*; D009865:Oocytes / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D012313:RNA / Q000032:analysis* / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000032:analysis*; D014981:Xenopus
chemical_list: D016601:RNA-Binding Proteins; D012313:RNA
keywords: ISH-PLA; RNA; protein-nucleic acid interactions