This detailed volume explores the continuing techniques ofstudying RNA-protein complexes and interactions as research in these areasexpand. After an introductory chapter,the book continues with ways to purify RNA-protein complexes assembled in cellsor in isolated cellular extracts, methods for measuring various biochemicalactivities of RNA-interacting proteins or ribonucleoproteins, biochemicalmethods for measuring direct RNA-protein contact, as well as various new orinnovative methods pertinent to the subject. Written for the highly successful Methods in Molecular Biology series,chapters…mehr
This detailed volume explores the continuing techniques ofstudying RNA-protein complexes and interactions as research in these areasexpand. After an introductory chapter,the book continues with ways to purify RNA-protein complexes assembled in cellsor in isolated cellular extracts, methods for measuring various biochemicalactivities of RNA-interacting proteins or ribonucleoproteins, biochemicalmethods for measuring direct RNA-protein contact, as well as various new orinnovative methods pertinent to the subject. Written for the highly successful Methods in Molecular Biology series,chapters contain brief introductions to their respective topics, lists of thenecessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, RNA-ProteinComplexes and Interactions: Methods and Protocols provides a set of usefulprotocols, both basic and advanced, designed to inspire researchers workingwith RNA and RNA-interacting proteins.
Characterization ofRNA-Protein Interactions: Lessons from Two RNA-Binding Proteins, SRSF1 andSRSF2.- Identification of mRNA-Interacting Factors by MS2-TRAP (MS2-Tagged RNAAffinity Purification).- Biotin-Streptavidin Affinity Purification ofRNA-Protein Complexes Assembled In Vitro.- Detecting RNA-Protein InteractionUsing End-Labeled Biotinylated RNA Oligonucleotides and Immunoblotting.- Purificationof RNA-Protein Splicing Complexes Using a Tagged Protein from In Vitro SplicingReaction Mixture.- Loading of Argonaute Protein with Small Duplex RNA inCellular Extracts.- In Vitro Analysis of Ribonucleoprotein Complex Remodelingand Disassembly.- Single-Turnover Kinetics of Methyl Transfer to tRNA byMethyltransferases.- Purification and Functional Reconstitution of Box H/ACARibonucleoprotein Particles.- Northwestern Blot Analysis: Detecting RNA-ProteinInteraction After Gel Separation of Protein Mixture.- Measuring mRNATranslation by Polysome Profiling.- Genome-Wide Profiling of RNA-ProteinInteractions Using CLIP-Seq.- Identification of Endogenous mRNA-BindingProteins in Yeast Using Crosslinking and PolyA Enrichment.- Ribo-ProteomicsApproach to Profile RNA-Protein and Protein-Protein Interaction Networks.- Detectionof Protein-Protein Interaction Within an RNA-Protein Complex ViaUnnatural-Amino-Acid-Mediated Photochemical Crosslinking.- Evolution ofCell-Type-Specific RNA Aptamers Via Live Cell-Based SELEX.- mCarts: Genome-WidePrediction of Clustered Sequence Motifs as Binding Sites for RNA BindingProteins.- Design of RNA-Binding Proteins: Manipulate Alternative Splicing inHuman Cells with Artificial Splicing Factors.- Informational Suppression toProbe RNA:RNA Interactions in the Context of Ribonucleoproteins: U1 and 5' Splice-SiteBase-Pairing.- Analysis of Alternative Pre-RNA Splicing in the Mouse RetinaUsing a Fluorescent Reporter.
Characterization ofRNA-Protein Interactions: Lessons from Two RNA-Binding Proteins, SRSF1 andSRSF2.- Identification of mRNA-Interacting Factors by MS2-TRAP (MS2-Tagged RNAAffinity Purification).- Biotin-Streptavidin Affinity Purification ofRNA-Protein Complexes Assembled In Vitro.- Detecting RNA-Protein InteractionUsing End-Labeled Biotinylated RNA Oligonucleotides and Immunoblotting.- Purificationof RNA-Protein Splicing Complexes Using a Tagged Protein from In Vitro SplicingReaction Mixture.- Loading of Argonaute Protein with Small Duplex RNA inCellular Extracts.- In Vitro Analysis of Ribonucleoprotein Complex Remodelingand Disassembly.- Single-Turnover Kinetics of Methyl Transfer to tRNA byMethyltransferases.- Purification and Functional Reconstitution of Box H/ACARibonucleoprotein Particles.- Northwestern Blot Analysis: Detecting RNA-ProteinInteraction After Gel Separation of Protein Mixture.- Measuring mRNATranslation by Polysome Profiling.- Genome-Wide Profiling of RNA-ProteinInteractions Using CLIP-Seq.- Identification of Endogenous mRNA-BindingProteins in Yeast Using Crosslinking and PolyA Enrichment.- Ribo-ProteomicsApproach to Profile RNA-Protein and Protein-Protein Interaction Networks.- Detectionof Protein-Protein Interaction Within an RNA-Protein Complex ViaUnnatural-Amino-Acid-Mediated Photochemical Crosslinking.- Evolution ofCell-Type-Specific RNA Aptamers Via Live Cell-Based SELEX.- mCarts: Genome-WidePrediction of Clustered Sequence Motifs as Binding Sites for RNA BindingProteins.- Design of RNA-Binding Proteins: Manipulate Alternative Splicing inHuman Cells with Artificial Splicing Factors.- Informational Suppression toProbe RNA:RNA Interactions in the Context of Ribonucleoproteins: U1 and 5' Splice-SiteBase-Pairing.- Analysis of Alternative Pre-RNA Splicing in the Mouse RetinaUsing a Fluorescent Reporter.
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