Full Description
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.
Contents
WT1 Mutation in Childhood Cancer.- WT1 and Renal Disease.- The Role of WT1 in Embryonic Development and Normal Organ Homeostasis.- Tools and Techniques for WT1-Based Lineage Tracing.- Biological Systems and Methods for Studying WT1 in the Epicardium.- Isolation and Colony Formation of Murine Bone and Bone Marrow Cells.- Isolation and Fluorescent Activated Cell Sorting of Murine WT1-Expressing Adipocyte Precursor Cells.- In Vivo Assays for Assessing the Role of the Wilms' Tumor Suppressor 1 (WT1) in Angiogenesis.- Multiphoton Microscopy for Visualizing Lipids in Tissue.- Function and Regulation of the Wilm's Tumor Suppressor 1 (WT1) Gene in Fish.- Immunofluorescence Staining of WT1 on Sections of Zebrafish Embryos and Larvae.- Fluorescence Activated Cell Sorting (FACS) Protocol for Podocyte Isolation in Adult Zebrafish.- In Vitro Transcription to Study WT1 Function.- Measuring Equilibrium Binding Constants for the WT1-DNAInteraction Using a Filter Binding Assay.- Identifying Direct Downstream Targets: WT1 ChIP-Seq Analysis.- WT1 Associated Protein-Protein Interaction Networks.- Methods to Identify and Validate WT1-RNA Interaction.- Bioinformatic Analysis of Next Generation Sequencing Data to Identify WT1 Associated Differential Gene and Isoform Expression.- Immunotherapy Targeting WT1: Designing a Protocol for WT1 Peptide-Based Cancer Vaccine.
