The Wilms' Tumor (Wt1) Gene: Methods and Protocols

ISBN-13: 9781493940219 / Angielski / Twarda / 2016 / 235 str.

Nicholas Hastie

Spis treści

1. WT1 Mutation in Childhood Cancer
Jocelyn Charlton and Kathy Pritchard-Jones

2. WT1 and Renal Disease
Eve Miller-Hodges

3. The Role of WT1 in Embryonic Development and Normal Organ Homeostasis
Bettina Wilm and Ramon Muñoz-Chapuli

4. Tools and Techniques for WT1-Based Lineage Tracing
Bettina Wilm and and Ramon Muñoz-Chapuli

5. Biological Systems and Methods for Studying WT1 in the Epicardium
Victor Velecela and Ofelia M. Martínez Estrada

6. Isolation and Colony Formation of Murine Bone and Bone Marrow Cells
Sophie McHaffie and You-Yong Chau

7. Isolation and Fluorescent Activated Cell Sorting of Murine WT1-Expressing Adipocyte Precursor  Cells
Louise Cleal and YouYing Chau

8. In Vivo Assays for Assessing the Role of the Wilms’ Tumor Suppressor 1 (WT1) in Angiogenesis
Richard McGregor, R. Ogley, Patrick Hadoke, and Nicholas Hastie

9. Multiphoton Microscopy for Visualizing Lipids in Tissue
Martin Lee and Alan Serrels

10. Function and Regulation of the Wilm’s Tumor Suppressor 1 (WT1) Gene in Fish
Birgit Perner, Thomas J. D. Bates, Uta Naumann, and Christoph Englert

11. Immunofluorescence Staining of WT1 on Sections of Zebrafish Embryos and Larvae
Birgit Perner and Christoph Englert

12. Fluorecence Activated Cell Sorting (FACS) Protocol for Podocyte Isolation in Adult Zebrafish
Thomas J. D. Bates, Uta Naumann, and Christoph Englert

13. In Vitro Transcription to Study WT1 Function
Stefan G. E. Roberts

14. Measuring Equilibrium Binding Constants for the WT1-DNA Interaction Using a Filter Binding Assay
Paul J. Romaniuk

15. Identifying Direct Downstream Targets: WT1 ChIP-Seq Analysis
Fabio da Silva, Filippo Massa, and Andreas Schedl

16. WT1 Associated Protein-Protein Interaction Networks
Ruthrothaselvi Bharathavikru and Alex von Kriegsheim

17. Methods to Identify and Validate WT1-RNA Interaction
Ruthrothaselvi Bharathavikru and Tatiana Dudnakova

18. Bioinformatic Analysis of Next Generation Sequencing Data to Identify WT1 Associated Differential Gene and Isoform Expression
Stuart Aitken and Ruthrothaselvi Bharathavikru

19. Immunotherapy Targeting WT1: Designing a Protocol for WT1 Peptide-Based Cancer Vaccine
Sumiyuki Nishida and Haruo Sugiyama                      

Blurb

This volume provides a thorough overview of the Wilms’ Tumour Gene (WT1). The book begins with three review chapters that cover the involvement of WT1 in pediatric cancer, kidney disease, and tissue development and homeostasis. The next few chapters discuss cell marking and lineage tracing, epicardial cell methodology, colony forming assays for bone marrow stem cells, angiogenesis assays and zebrafish tools.  The next group of chapters explores the latest tools in genomics, molecular biology, and biochemistry. They discuss dissecting transcription factor function in cell free systems, ChiP seq, proteomics, RNA interactome, and multiphoton imaging of lipids, measuring the binding constants of protein-nucleic acid interactions, and bioinformatics approaches for analyzing Next Generation Sequence data. The final chapter discusses protocols for clinical trials for immune therapy using anti-WT1 peptides. 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.

Practical and thorough, The Wilms’ Tumour (WT1) Gene: Methods and Protocols is a valuable resource for anyone who is interested in the diverse methodologies used in WT1 research.