title: Single Molecule Approaches in RNA-Protein Interactions.
abstract: RNA-protein interactions govern every aspect of RNA metabolism, and aberrant RNA-binding proteins are the cause of hundreds of genetic diseases. Quantitative measurements of these interactions are necessary in order to understand mechanisms leading to diseases and to develop efficient therapies. Existing methods of RNA-protein interactome capture can afford a comprehensive snapshot of RNA-protein interaction networks but lack the ability to characterize the dynamics of these interactions. As all ensemble methods, their resolution is also limited by statistical averaging. Here we discuss recent advances in single molecule techniques that have the potential to tackle these challenges. We also provide a thorough overview of single molecule colocalization microscopy and the essential protein and RNA tagging and detection techniques.
authors.affiliation: Howard Hughes Medical Institute, University of Massachusetts Medical School, Worcester, MA, USA.
authors.forename: Victor
authors.initials: V
authors.lastname: Serebrov
authors.affiliation: Howard Hughes Medical Institute, University of Massachusetts Medical School, Worcester, MA, USA. melissa.moore@umassmed.edu.
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Moore
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis; D006801:Humans; D006867:Hydrolases / Q000737:chemistry; D008164:Luminescent Proteins / Q000032:analysis; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D000072760:Single Molecule Imaging / Q000295:instrumentation / Q000379:methods*; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D010446:Peptide Fragments; D016601:RNA-Binding Proteins; D012313:RNA; D013762:Tetrahydrofolate Dehydrogenase; D019853:O(6)-Methylguanine-DNA Methyltransferase; D006867:Hydrolases; C046137:haloalkane dehalogenase
keywords: CoSMoS; HILO; Protein tags; Single molecule imaging; TIRF