Main description:

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.

Contents:

Preface...Dedication...Table of Contents...Contributing Authors...
Part I Imaging RNAs and Gene Expression in Cells and Organisms

1) Imaging Single mRNA Molecules in Mammalian Cells using an Optimized MS2-MCP SystemMaria Vera, Evelina Tutucci, and Robert H. Singer
2) Live-Cell Imaging of Long Non-Coding RNAs using Molecular BeaconsYachen Ying, Shiqi Mao, Christopher J. Krueger, and Antony K. Chen
3) Real-Time Observation of Localization and EXpression (ROLEX) System for Live Imaging of the Transcriptional Activity and Nuclear Position of a Specific Endogenous GeneHiroshi Ochiai
4) Visualization of Single mRNAs in Live NeuronsJae Youn Shim, Byung Hun Lee, and Hye Yoon Park
5) Lightening Up Gene Activation in Living Drosophila EmbryosCarola Fernandez and Mounia Lagha
6) Quantification of mRNA Turnover in Living Cells: A Pipeline for TREAT Data AnalysisFranka Voigt, Jan Eglinger, and Jeffery A. Chao
7) Imaging rRNA Methylation in Bacteria by MR-FISHKristina A. Ganzinger, Martin R. Challand, James Spencer, David Klenerman, and Rohan T. Ranasinghe
8) Detection of mRNA Transfer between Mammalian Cells in Co-Culture by Single Molecule Fluorescent In Situ Hybridization (smFISH)Gal Haimovich and Jeffrey E. Gerst
9) Live-Cell Imaging of mRNP-NPC Interactions in Budding YeastAzra Lari, Farzin Farzam, Pierre Bensidoun, Marlene Oeffing, Daniel Zenklusen, David Grunwald, and Ben Montpetit
10) Detection of mRNAs Anchored to the Nuclear Envelope during Export Inhibition in Living CellsRakefet Ben-Yishay and Yaron Shav-Tal
Part II Imaging Chromosomes, Genes, and Transcription Factors
11) Programmable Chromosome Painting with OligopaintsSon C. Nguyen and Eric F. Joyce
12) Imaging Chromosome Territory and Gene Loci Positions in Cells Grown on Soft Matrices Roopali Pradhan and Kundan Sengupta
13) Targeted Manipulation/Repositioning of Subcellular Structures and MoleculesKathrin S. Heinz and M. Cristina Cardoso
14) Visualization of Endogenous Transcription Factors in Single Cells using an Antibody Electroporation-Based Imaging ApproachSascha Conic, Dominique Desplancq, Alexia Ferrand, Nacho Molina, Etienne Weiss, and Laszlo Tora
15) Monitoring Transcription Factor Oligomerization in Single Living Cells by Numbers and Brightness AnalysisEugenia Cammarota and Davide Mazza
16) Visualizing Transcription Factor Binding on Mitotic Chromosomes using Single Molecule Live-Cell Imaging James Z.J. Kwan, Thomas F. Nguyen, and Sheila S. Teves
17) Light-Induced Transcription Activation for Time-Lapse Microscopy Experiments in Living CellsJorge Trojanowski, Anne Rademacher, Fabian Erdel, and Karsten Rippe