RNA Chaperones

This second edition provides new and up-to-date methods to study RNA chaperones in vitro and protocols for cell-based assays. Chapter guide readers through bacterial proteins, proteins from eukaryotic cells, complex interactions between RNA chaperones, and the folding and unfolding of proteins. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, RNA Chaperones: Methods and Protocols, Second Edition aims to be comprehensive guide for researchers in the field

A two-step purification strategy for RNA and RNA-Protein complexes for crystallization. - Analysis of tRNA aminoacylation by northern blotting and multiplex small RNA-Seq. - Characterization of RNA Structure Dynamics with trans-factors using Single-Molecule Optical Tweezers. - Specific nucleic acid chaperone activity of HIV-1 nucleocapsid protein deduced from hairpin unfolding. - A dual-color electrophoretic mobility shift assay to examine RNA binding and RNA chaperone activity simultaneously. - Use of tRNA mediated suppression to assess RNA chaperone function. - Salt-dependent modulation of the RNA chaperone activity of RNA-binding protein La. - Kinetic and thermodynamic analyses of RNA protein interactions. - Detection of MicroRNAs released from Argonautes. - Mapping the RNA chaperone activity of the T. brucei editosome using SHAPE-chemical probing. - Using immunoprecipitation followed by dot blotting to identify oligonucleotide-protein interactions in cells. - In vivo RNA chemical footprinting analysis in Archaea. - RNA structure analysis by chemical probing with DMS and CMCT. - Prediction of intrinsically disordered regions that bind RNA with DisoRDPbind. - Delivering Molecular Beacons via an Electroporation-Based Approach Enables Live-Cell Imaging of Single RNA Transcripts and Genomic Loci. - Site-specific dual-color labeling of long RNAs. - Characterizing complex nucleic acid interactions of LINE1 ORF1p by single molecule force spectroscopy. - Isolation and analysis of bacterial ribosomes through sucrose gradient ultracentrifugation.