title: UV cross-linking of interacting RNA and protein in cultured cells.
abstract: RNA-protein interactions play indispensable roles in the regulation of cellular functions. Biochemical characterization of these complexes is often done by immunoprecipitation (IP) of RNA-binding proteins (RBPs) followed by identification of co-immunoprecipitated RNAs. This protocol couples ultraviolet (UV) irradiation with IP to determine whether a specific protein interacts directly with a specific RNA in living cells.
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX, USA.
authors.forename: Emi
authors.initials: E
authors.lastname: Sei
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX, USA. Electronic address: nicholas.conrad@utsouthwestern.edu.
authors.forename: Nicholas K
authors.initials: NK
authors.lastname: Conrad
mesh_terms: D015152:Blotting, Northern; D002478:Cells, Cultured; D047468:Immunoprecipitation; D011485:Protein Binding; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D016601:RNA-Binding Proteins; D012313:RNA
keywords: Cross-linking protocol (CLIP); Immunoprecipitation; Northern blotting; Proteinase K treatment; RNA immunoprecipitation (RIP) and cell-mixing protocols; RNA–protein interactions; Ribonucleoprotein particles (RNPs); Sodium dodecyl sulfate (SDS) Lysis Buffer; UV cross-linking; Western blotting