Imaging Gene Expression: Methods and Protocols

ISBN-13: 9781627035255 / Angielski / Twarda / 2013 / 368 str.

Yaron Shav-Tal

Opis

As imaging technologies and approaches have evolved, the scope of certain imaging techniques has moved far beyond the production of purely illustrative images or appealing time-lapse movies to providing the scientist with a rich range of ways to measure and quantify the biological process and outcome of gene expression. In Imaging Gene Expression: Methods and Protocols, expert authors offer up-to-date approaches and protocols that scientists in the field have developed, which would benefit the broader scientific community. Divided in three convenient parts, this detailed book covers the output of a gene, namely the RNA molecules that are transcribed from the gene and the way by which these molecules can be tracked or quantified in fixed or living cells, protocols that focus on the gene, DNA, or chromatin, as well as a variety of ways by which nuclear processes intertwined with gene expression can be followed and quantified in living cells as well as approaches for studying several sub-nuclear structures found in eukaryotic cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective subjects, lists of materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and up-to-date, Imaging Gene Expression: Methods and Protocols will serve researchers working toward imaging in the context of complete organisms.

Spis treści

Part I: Imaging Gene Expression and RNA Dynamics in Cells and Organisms

 

1. High-Throughput Fluorescence-Based Screen to Identify Factors Involved in Nuclear Receptor Recruitment to Response Elements

            Tina B. Miranda, Ty C. Voss, and Gordon L. Hager

 

2. Live-Cell Imaging Combined with Immunofluorescence, RNA, or DNA FISH to Study the Nuclear Dynamics and Expression of the X-Inactivation Center

            Tim Pollex, Tristan Piolot, and Edith Heard

 

3. Single Molecule Resolution Fluorescent In Situ Hybridization (smFISH) in the Yeast S. cerevisiae

            Samir Rahman and Daniel Zenklusen

 

4. Measuring Transcription Dynamics in Living Cells Using Fluctuation Analysis

            Matthew L. Ferguson and Daniel R. Larson

 

5. Tracking Nuclear Poly(A) RNA Movement Within and Among Speckle Nuclear Bodies and the Surrounding Nucleoplasm

            Joan C. Ritland Politz and Thoru Pederson

 

6. Nuclear Trafficking and Export of Single, Native mRNPs in Chironomus tentans Salivary Gland Cells

            Tim P. Kaminski, Jan-Hendrik Spille, Claudio Nietzel, Jan Peter Siebrasse, and Ulrich Kubitscheck

 

7. Single mRNP Tracking in Living Mammalian Cells

            Alon Kalo, Pinhas Kafri, and Yaron Shav-Tal

 

8. Imaging Nascent RNA Dynamics in Dictyostelium

            Jonathan R. Chubb, Michelle Stevense, Danielle Cannon, Tetsuya Muramoto, and Adam Corrigan

 

 

Part II: Imaging the Genome and Chromatin Dynamics

 

9. Monitoring Dynamic Binding of Chromatin Proteins In Vivo by Single Molecule Tracking

            Davide Mazza, Sourav Ganguly, and James G. McNally

 

10. Single Particle Tracking for Studying the Dynamic Properties of Genomic Regions in Live Cells

            Irena Bronshtein Berger, Eldad Kepten, and Yuval Garini

 

11. Microscopic Analysis of Chromatin Localization and Dynamics in C. elegans

            Christian Lanctôt and Peter Meister

 

12. Measuring the Dynamics of Chromatin Proteins During Differentiation

            Arigela Harikumar and Eran Meshorer

 

13. Electron Spectroscopic Tomography of Specific Chromatin Domains

            Liron Even-Faitelson, Eden Fussner, Ren Li, Mike Strauss, and David P. Bazett-Jones

 

14. BAC Manipulations for Making BAC Transgene Arrays

            Nimish Khanna, Qian Bian, Matt Plutz, and Andrew S. Belmont

 

 

Part III: Imaging Nuclear Processes and Sub-Nuclear Structures

 

15. Spatio-Temporal Visualization of DNA Replication Dynamics

            Marius Reinhart, Corella S. Casas-Delucchi, and M. Cristina Cardoso

 

16. The Dynamics of DNA Damage Repair and Transcription

            Niraj M. Shanbhag and Roger A. Greenberg

 

17. Fluorescence Microscopy-Based High-Throughput Screening for Factors Involved in Gene Silencing

            Sebastian Bultmann and Heinrich Leonhardt

18. Actin as a Model for the Study of Nucleocytoplasmic Shuttling and Nuclear Dynamics

            Kari-Pekka Skarp and Maria K. Vartiainen

 

19. Isochronal Visualization of Transcription and Proteasomal Proteolysis in Cell Culture or in the Model Organism, Caenorhabditis elegan

            Anna von Mikecz and Andrea Scharf

 

20. Considering Discrete Protein Pools When Measuring the Dynamics of Nuclear Membrane Proteins

            Nikolaj Zuleger, David A. Kelly, and Eric C. Schirmer

 

21. Correlative Microscopy of Individual Cells: Sequential Application of Microscopic Systems with Increasing Resolution to Study the Nuclear Landscape

            Barbara Hübner, Thomas Cremer, and Jürgen Neumann

 

22. Time-Lapse, Photo-Activation, and Photo-Bleaching Imaging of Nucleolar Assembly after Mitosis

            Danièle Hernandez-Verdun, Emilie Louvet, and Eleonora Muro

 

23. Nucleation of Nuclear Bodies

            Miroslav Dundr