Ewing Sarcoma
Description
This second edition volume aims at providing new and updated methods detailing the most effective and widely employed protocols used in the Ewing sarcoma field. Chapters guide the readers through molecular biology techniques used to analyze Ewing sarcoma including tissue preservation and methods used in morpho-molecular diagnostics and routine immune-histological analysis. Additional wet-lab protocols comprise i n vitro techniques including re-expression and functional analyses of target genes together with in vivo tools and models such as patient-derived xenografts (PDX) and orthotopic xenografts for metastasis assessment in Ewing sarcoma.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 clearly written, Ewing sarcoma: Methods and Protocols, Second Edition aims to provide methods to further the study of Ewing sarcoma and bring researchers one step forward in the quest to fight this devastating disease.
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Content
Germline variation and somatic alterations in Ewing sarcoma. - Western blot analysis in Ewing sarcoma.- Tissue preservation and FFPE samples: Optimized nucleic acids isolation in Ewing sarcoma.Untargeted metabolite screening of Ewing sarcoma cells - and tissues with Liquid Chromatography - Tandem Mass Spectrometry (LC-MS/MS).- (Immuno-)histological analysis of Ewing sarcoma.- Molecular approaches to diagnosis diagnosis in Ewing sarcoma: RT-PCR.- Molecular approaches to diagnosis in Ewing sarcoma: Targeted RNA sequencing.- Ewing-specific (re)expression models.- Analysis of regulatory sequences by dual-luciferase reporter assays in Ewing sarcoma.- Proliferation assessment by Trypan blue exclusion in Ewing sarcoma.- of resazurin and ATP detection cell viability assays for drug sensitivity screening in Ewing sarcoma.- Analysis of migration and invasion in Ewing sarcoma.- Analysis of colony formation capacity in Ewing sarcoma.- Genetic and epigenetic manipulation using CRISPR techniques in Ewing sarcoma.- Post-translational regulation using degrons in Ewing sarcoma.- 3D Ewing sarcoma modelling for metabolomic studies.- Genetically engineered mouse model in Ewing sarcoma.- Tumor growth analysis of Ewing sarcoma cell lines using subcutaneous xenografts in mice.- Metastasis assessment in Ewing sarcoma using orthotopic xenografts.- Orthotopic implants in mice.- Ewing sarcoma PDX models.- Using zebrafish larvae as a xenotransplantation model to study Ewing sarcoma.- Chorioallantoic membrane assay to assess primary and metastatic tumor growth in Ewing sarcoma.- A Drosophila study model for EWS::FLI1-induced molecular pathway- A practical compass to biological digital resources and omics integration in Ewing sarcoma. - Chromatin immunoprecipitation followed by next-generation sequencing in Ewing sarcoma.- Epigenetic analysis in Ewing sarcoma.- Leveraging single-cell RNA sequencing to characterize murine Ewing sarcoma xenografts.- Spatial transcriptomics analysis of Ewing sarcoma.