Cell Cycle Oscillators: Methods and Protocols

ISBN-13: 9781071615379 / Angielski / Twarda / 2021 / 340 str.

Amanda S. Coutts; Louise Weston

Spis treści

1. Cell Cycle Control: A System of Interlinking Oscillators

            Randy Y.C. Poon

 

2. Substrate Phosphorylation Rates as an In Vivo Measurement of Kinase Activity

            Matthew P. Swaffer

 

3. Real Time Monitoring of APC/C-Mediated Substrate Degradation Using Xenopus laevis Egg Extracts

            Julia Kamenz, Renping Qiao, Qiong Yang, and James E. Ferrell, Jr.

 

4. Fluorescent Peptide Biosensors for Probing CDK Kinase Activity in Cell Extracts

            Morgan Pellerano and May C. Morris

 

5. Phosphatase and Kinase Substrate Specificity Profiling with Pooled Synthetic Peptides and Mass Spectrometry

            Andrew G. DeMarco, Pete E. Pascuzzi, W. Andy Tao, and Mark C. Hall

 

6. Whole-Mount Immunostaining for the Identification of Histone Modifications in the S-Phase Nuclei of Arabidopsis Roots

            Hirotomo Takatsuka and Masaaki Umeda

 

7. Construction of a Full-Length 3’UTR Reporter System for Identification of Cell-Cycle Regulating MicroRNAs

            Dominika Kazmierczak and Per Hydbring

 

8. Purification of Cyclin-Dependent Kinase Fusion Complexes for In Vitro Analysis

            Mardo Kõivomägi

 

9. Optimizing Cell Synchronization Using Nocodazole or Double Thymidine Block

            Arif A. Surani, Sergio L. Colombo, George Barlow, Gemma A. Foulds, and Cristina Montiel-Duarte

 

10. Highly Synchronous Mitotic Progression in Schizosaccharomyces pombe Upon Relief of Transient Cdc2-asM17 Inhibition

            Pawan Singh, Lenka Halova, and Iain Michael Hagan

 

11. Elucidating Human Mitosis Using an Anaphase-Like Cell-Free System

            Danit Wasserman, Sapir Nachum, Meirav Noach-Hirsh, Naomi Auerbach, Evelin Sheinberger-Chorni, Taylor P Enrico, Roxane Lahmi, Michael J. Emanuele, and Amit Tzur

 

12. EDU (5-Ethynyl-2′-Deoxyuridine)-Coupled-Fluorescence-Intensity Analysis: Determining Absolute Parameters of the Cell Cycle

            João A. Ferreira, Marco Neves, Miguel Alpalhão, Pedro Pereira, Daniela Cunha, Fernando Ferreira, René Santus, Ana E. Sousa, and Paulo L. Filipe

 

13. High-Resolution Analysis of Centrosome Behavior During Mitosis

            Vanessa Nunes, Margarida Dantas, Joana T. Lima, and Jorge G. Ferreira

 

14. Assaying Cell Cycle Progression via Flow Cytometry in CRISPR/Cas9-Treated Cells

            Jonathan M. Geisinger and Tim Stearns

 

15. Use of Mitotic Protein Kinase Inhibitors and Phospho-Specific Antibodies to Monitor Protein Phosphorylation During the Cell Cycle

            Isha Nasa, Greg B. Moorhead, and Arminja N. Kettenbach

 

16. Visualization of Radiation-Induced Cell Cycle Kinetics with a Fluorescent Ubiquitination-Based Cell Cycle Indicator (Fucci)

            Atsushi Kaida and Masahiko Miura

 

17. Dynamic Behavior of Inactive X Chromosome Territory During the Cell Cycle as Revealed by H3K27me3-Specific Intracellular Antibody

            Yuko Sato and Hiroshi Kimura

 

18. Analyzing Centrioles and Cilia by Expansion Microscopy

            Dong Kong and Jadranka Loncarek

 

19. Analysis of Cell Cycle Progression in the Budding Yeast S. cerevisiae

            Deniz Pirincci Ercan and Frank Uhlmann

 

20. Application of PALM Super Resolution Microscopy to the Analysis of Microtubule-Organizing Centers (MTOCs) in Aspergillus nidulans

            Xiaolei Gao, Reinhard Fischer, and Norio Takeshita

 

21. Live Imaging and Analysis of Cilia and Cell Cycle Dynamics with the Arl13bCerulean-Fucci2a Biosensor and Fucci Tools

            Melinda Van Kerckvoorde, Matthew J. Ford, Patricia L. Yeyati, Pleasantine Mill, and Richard L. Mort

 

22. Calorimetric Heat Dissipation Measurements of Developing Zebrafish Embryos

            Jonathan Rodenfels and Karla M. Neugebauer

 

23. The Conditional Knockout Analogous System: CRISPR-Mediated Knock Out Together with Inducible Degron and Transcription-Controlled Expression

            Hoi Tang Ma

Blurb

This book brings together a unique collection of protocols that cover novel and specialized techniques as well as updated and improved adaptations of more standard procedures involving the cell cycle and its regulation by oscillatory networks, exploring recent progress in the field from both holistic and reductionist perspectives. The edition provides a space for researchers to highlight and explore the latest developments in molecular biology and biochemical techniques for studying oscillatory networks and to share these across the research community to facilitate further progress. 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. 

Authoritative and up-to-date, Cell Cycle Oscillators: Methods and Protocols, Second Edition serves as an invaluable guide for those new to the field as well as the more experienced scientist.

Chapters 19 and 21 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.