Imaging Gene Expression: Methods and Protocols

ISBN-13: 9781493996766 / Angielski / Miękka / 2020 / 272 str.

Yaron Shav-Tal

Opis

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.

Spis treści

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 System

Maria Vera, Evelina Tutucci, and Robert H. Singer

2) Live-Cell Imaging of Long Non-Coding RNAs using Molecular Beacons

Yachen 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 Gene

Hiroshi Ochiai

4) Visualization of Single mRNAs in Live Neurons

Jae Youn Shim, Byung Hun Lee, and Hye Yoon Park

5) Lightening Up Gene Activation in Living Drosophila Embryos

Carola Fernandez and Mounia Lagha

6) Quantification of mRNA Turnover in Living Cells: A Pipeline for TREAT Data Analysis

Franka Voigt, Jan Eglinger, and Jeffery A. Chao

7) Imaging rRNA Methylation in Bacteria by MR-FISH

Kristina 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 Yeast

Azra 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 Cells

Rakefet Ben-Yishay and Yaron Shav-Tal

Part II Imaging Chromosomes, Genes, and Transcription Factors

11) Programmable Chromosome Painting with Oligopaints

Son 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 Molecules

Kathrin S. Heinz and M. Cristina Cardoso

14) Visualization of Endogenous Transcription Factors in Single Cells using an Antibody Electroporation-Based Imaging Approach

Sascha 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 Analysis

Eugenia 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 Cells

Jorge Trojanowski, Anne Rademacher, Fabian Erdel, and Karsten Rippe