This book provides a wide spectrum of methods to study RNA chaperones in vitro , at the single molecule level, and protocols useful for cell-based assays. Beginning with a section on a number of bacterial proteins for study, the volume also explores proteins from eukaryotic cells and how to delve into the 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, RNA Chaperones: Methods and Protocols serves as an ideal guide for scientists and students interested in RNA biology and RNA chaperones.

Chapter 3 is available Open Access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Klappentext

This book provides a wide spectrum of methods to study RNA chaperones in vitro, at the single molecule level, and protocols useful for cell-based assays. Beginning with a section on a number of bacterial proteins for study, the volume also explores proteins from eukaryotic cells and how to delve into the 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, RNA Chaperones: Methods and Protocols serves as an ideal guide for scientists and students interested in RNA biology and RNA chaperones. Chapter 3 is available Open Access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Inhalt

Biochemical Methods for the Study of the FinO Family of Bacterial RNA Chaperones.- Quantitative Analysis of RNA Chaperone Activity by Native Gel Electrophoresis and Fluorescence Spectroscopy.- Fluorescent Molecular Beacons Mimicking RNA Secondary Structures to Study RNA Chaperone Activity.- Specific Nucleic Acid Chaperone Activity of HIV-1 Nucleocapsid Protein Deduced from Hairpin Unfolding.- Real-Time Fluorescence-Based Approaches to Disentangle Mechanisms of a Protein's RNA Chaperone Activity.- 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 RNAProtein Interactions.- Detection of MicroRNAs Released from Argonautes.- Mapping the RNA Chaperone Activity of the T. brucei Editosome Using SHAPE-Chemical Probing.- RNA Remodeling by RNA Chaperones Monitored by RNA Structure Probing.- In Vivo RNA Chemical Footprinting Analysis in Archaea.- RNA Structure Analysis by Chemical Probing with DMS and CMCT.- Disordered RNA Binding Region Prediction 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.- Single-Molecule FRET Assay for Studying Cotranscriptional RNA Folding.- Characterizing Complex Nucleic Acid Interactions of LINE1 ORF1p by Single Molecule Force Spectroscopy.- Isolation and Analysis of Bacterial Ribosomes through Sucrose Gradient Ultracentrifugation.