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This volume compiles methodologies used for detailed studies of histone variants, from their basic properties to their functional roles in chromatin and as vectors of epigenetic information. Its four sections cover experimental approaches to probe the biochemistry of histone variants and variant nucleosomes; their dynamics throughout the cell cycle and at specific genomic locations; their functional impact on chromosome organization and genome stability; and their importance in development, disease and evolution. Written in the highly successful Methods in Molecular Biology series format, chapters include lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, illustrations on the expected results, and tips on troubleshooting and avoiding known pitfalls. This anthology brings together contributions from many of the most highly recognized investigators in this community.
Cutting-edge and thorough, Histone Variants: Methods and Protocols is a valuable resource for anyone interested in exploring the experimental and collaborative possibilities in this evolving field.
List of contents
Methods for Preparing Nucleosomes Containing Histone Variants.- Characterization of Post-Translational Modifications on Histone Variants.- Purification of Histone Variant-Interacting Chaperone Complexes.- Detection of Histone Modification Dynamics during the Cell Cycle by MS-Based Proteomics.- Histone Native Chromatin Immunprecipitation.- How to Tackle Challenging ChIP-seq, with Long-Range Cross-Linking, using ATRX as an Example.- Time-ChIP : A Method to Determine Long-Term Locus-Specific Nucleosome Inheritance.- MINCE-seq : Mapping In Vivo Nascent Chromatin with EdU and Sequencing.- RChIP-seq : Chromatin-Associated RNA Sequencing in Developmentally Staged Mouse Testes.- Bioinformatics Analysis of Nucleosome and Histone Variant Positioning.- Imaging newly Synthesized and Old Histone Variant Dynamics Dependent on Chaperones Using the SNAP-Tag System.- Real Time de novo Deposition of Centromeric Histone-Associated Proteins Usingthe Auxin Inducible Degradation System.- Live Imaging of Parental Histone Variant Dynamics in UVC-Damaged Chromatin.- CRISPR/Cas9 Gene Editing of Human Histone H2A Variant H2AX and MacroH2A.- Studying the Evolution of Histone Variants Using Phylogeny.- Characterization of Post-Meiotic Male Germ Cell Genome Organizational States.- An Animal Model for Genetic Analysis of Multi-Gene Families : Cloning and Transgenesis of Large Tandemly-Repeated Histone Gene Clusters.- Imaging and Quantitation of Assembly Dynamics of the Centromeric Histone H3 Variant CENP-A in Drosophila melanogaster Spermatocytes by Immunofluorescence and Fluorescence In-Situ Hybridization (Immuno-FISH).
Summary
This volume compiles methodologies used for detailed studies of histone variants, from their basic properties to their functional roles in chromatin and as vectors of epigenetic information. Its four sections cover experimental approaches to probe the biochemistry of histone variants and variant nucleosomes; their dynamics throughout the cell cycle and at specific genomic locations; their functional impact on chromosome organization and genome stability; and their importance in development, disease and evolution. Written in the highly successful Methods in Molecular Biology series format, chapters include lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, illustrations on the expected results, and tips on troubleshooting and avoiding known pitfalls. This anthology brings together contributions from many of the most highly recognized investigators in this community.
Cutting-edge and thorough, Histone Variants: Methods and Protocols is a valuable resource for anyone interested in exploring the experimental and collaborative possibilities in this evolving field.
