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This volume explores the latest updates on microscopy approaches and techniques used by scientists studying in the field of gene expression imaging. These updates cover the technical design of the experiments and the expected outcomes. The chapters in this book are divided into two parts: Part One looks at the output of a gene, in particular the RNA molecules that are copied from the gene itself; and Part Two focuses on chromosomes, chromatin, and factors that bind DNA. Written in the highly successful Methods in Molecular Biology series format, 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. Cutting-edge and thorough, Imaging Gene Expression: Methods and Protocols, Second Edition is a valuable resource for any researcher interested in learning more about this evolving and important field.
List of contents
Imaging Single mRNA Molecules in Mammalian Cells using an Optimized MS2-MCP System.- Live-Cell Imaging of Long Non-Coding RNAs using Molecular Beacons.- Real-Time Observation of Localization and EXpression (ROLEX) System for Live Imaging of the Transcriptional Activity and Nuclear Position of a Specific Endogenous Gene.- Visualization of Single mRNAs in Live Neurons.- Lightening Up Gene Activation in Living Drosophila Embryos.- Quantification of mRNA Turnover in Living Cells: A Pipeline for TREAT Data Analysis.- Imaging rRNA Methylation in Bacteria by MR-FISH.- Detection of mRNA Transfer between Mammalian Cells in Co-Culture by Single Molecule Fluorescent In Situ Hybridization (smFISH).- Live-Cell Imaging of mRNP-NPC Interactions in Budding Yeast.- Detection of mRNAs Anchored to the Nuclear Envelope during Export Inhibition in Living Cells.- Programmable Chromosome Painting with Oligopaints.- Imaging Chromosome Territory and Gene Loci Positions in Cells Grown on Soft Matrices.- Targeted Manipulation/Repositioning of Subcellular Structures and Molecules.- Visualization of Endogenous Transcription Factors in Single Cells using an Antibody Electroporation-Based Imaging Approach.- Monitoring Transcription Factor Oligomerization in Single Living Cells by Numbers and Brightness Analysis.- Visualizing Transcription Factor Binding on Mitotic Chromosomes using Single Molecule Live-Cell Imaging.- Light-Induced Transcription Activation for Time-Lapse Microscopy Experiments in Living Cells.
