Characterization of RNA-Protein Interactions : Lessons from Two RNA-Binding Proteins, SRSF1 and SRSF2 -- Identification of mRNA-Interacting Factors by MS2-TRAP (MS2-Tagged RNA Affinity Purification) -- Biotin-Streptavidin Affinity Purification of RNA-Protein Complexes Assembled In Vitro -- Detecting RNA-Protein Interaction Using End-Labeled Biotinylated RNA Oligonucleotides and Immunoblotting -- Purification of RNA-Protein Splicing Complexes Using a Tagged Protein from In Vitro Splicing Reaction Mixture -- Loading of Argonaute Protein with Small Duplex RNA in Cellular Extracts -- In Vitro Analysis of Ribonucleoprotein Complex Remodeling and Disassembly -- Single-Turnover Kinetics of Methyl Transfer to tRNA by Methyltransferases -- Purification and Functional Reconstitution of Box H/ACA Ribonucleoprotein Particles -- Northwestern Blot Analysis: Detecting RNA-Protein Interaction After Gel Separation of Protein Mixture -- Measuring mRNA Translation by Polysome Profiling -- Genome-Wide Profiling of RNA-Protein Interactions Using CLIP-Seq -- Identification of Endogenous mRNA-Binding Proteins in Yeast Using Crosslinking and PolyA Enrichment -- Ribo-Proteomics Approach to Profile RNA-Protein and Protein-Protein Interaction Networks -- Detection of Protein-Protein Interaction Within an RNA-Protein Complex Via Unnatural-Amino-Acid-Mediated Photochemical Crosslinking -- Evolution of Cell-Type-Specific RNA Aptamers Via Live Cell-Based SELEX -- mCarts: Genome-Wide Prediction of Clustered Sequence Motifs as Binding Sites for RNA Binding Proteins -- Design of RNA-Binding Proteins: Manipulate Alternative Splicing in Human Cells with Artificial Splicing Factors -- Informational Suppression to Probe RNA:RNA Interactions in the Context of Ribonucleoproteins: U1 and 5{u2019} Splice-Site Base-Pairing -- Analysis of Alternative Pre-RNA Splicing in the Mouse Retina Using a Fluorescent Reporter.
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This detailed volume explores the continuing techniques of studying RNA-protein complexes and interactions as research in these areas expand. After an introductory chapter, it continues with ways to purify RNA-protein complexes assembled in cells or in isolated cellular extracts, methods for measuring various biochemical activities of RNA-interacting proteins or ribonucleoproteins, biochemical methods for measuring direct RNA-protein contact, as well as various new or innovative methods pertinent to the subject--