图书简介
This fully updated edition provides a series of methods for how best to assess functions of histone deacetylases and acetyltransferases. The disease-relevance of dysregulated protein deacetylation by overexpressed or aberrantly activated histone deacetylases has spurred an intense search for novel and improved inhibitors of these enzymes, as reflected in this collection. Expert contributors explore the generation and evaluation of novel histone deacetylase inhibitors and new and improved techniques to assess acetylation-dependent molecular mechanisms in vitro and in vivo. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, HDAC/HAT Function Assessment and Inhibitor Development: Methods and Protocols, Second Edition serves as an ideal guide for researchers seeking to further elucidate this vital area of study.
Analyzing Lymphoma Development and Progression Using HDACi in Mouse Models.- Colony Formation Assay to Test the Impact of HDACi on Leukemic Cells.- Defined Human Leukemic CD34 Liquid Cultures to Study HDAC/Transcriptional Repressor Complexes.- Development of a Cellular Model Mimicking Specific HDAC Inhibitors.- Evaluation of Anti-Tumor and On-Target Activity of HDAC Inhibitors with the Zebrafish Embryo Xenograft Model.- Human Platelet Lysate as Valid Cell Growth Additive to Assess Protein Acetylation.- Investigating Physiopathological Roles for Sirtuins in a Mouse Model.- Patient-Derived Organoids for In Vivo Validation of In Vitro Data.- Analysis of HDACi-Coupled Nanoparticles: Opportunities and Challenges.- Development of Pyrazine-Anilinobenzamides as Histone Deacetylase HDAC1-3 Selective Inhibitors and Biological Testing Against Pancreas Cancer Cell Lines.- Evaluation of Small Molecule HDAC Inhibitors through In Vitro and In Cellulo Approaches.- Synthesis, Biochemical, and Cellular Evaluation of HDAC6 Targeting Proteolysis Targeting Chimeras.- HDACi Delivery Systems Based on Cellulose Valproate Nanoparticles.- Synthesis and Characterization of Reversible Covalent HDAC4 Inhibitors.- Analysis of Cell Cycle and DNA Compaction Dependent Subnuclear Distribution of Histone Marks.- Analyzing the Effects of HDAC Inhibitors on DNA Damage and Associated Cytotoxicity in Primary Hepatocytes.- Assessment of HDAC Inhibitor-Induced Endoplasmic Reticulum (ER) Stress.- Assessment of Mitochondrial Dysfunctions after Sirtuin Inhibition.- Assessing the Effect of Histone Deacetylase Inhibitors on DNA Double-Strand Break Repair by Non-Homologous End Joining.- Assessing SIRT7 Activity In Vivo and In Vitro in Response to DNA Damage.- Methods to Evaluate the Effects of HAT/KAT Inhibition on SIAH2-Driven Reactive Oxygen Species Generation in Helicobacter pylori-Infected Gastric Epithelial Cells.- Monitoring Changes in Intracellular Reactive Oxygen Species Levels in Response to Histone Deacetylase Inhibitors.- Single-Cell Analysis of Histone Acetylation Dynamics at Replication Forks Using PLA and SIRF.- Testing the Effect of Histone Acetyltransferases on Local Chromatin Compaction.- A Genome-Wide CRISPR-Cas9 Loss-of-Function Screening to Identify Host Restriction Factors Modulating Oncolytic Virotherapy.- Cloning Strategy for HDAC1/HDAC2 Hybrid Protein Expression in Mammalian Cells.- Continuous Histone Deacylase Activity Assays.- CRISPR Activation/Interference Screen to Identify Genetic Networks in HDAC-Inhibitor Resistant Cells.- Determining Potency of Inhibitors Targeting Histone Deacetylase 6 by Quantification of Acetylated Tubulin in Cells.- Expression and Crystallization of HDAC6 Tandem Catalytic Domains.- Fluorescence Polarization-Based Competition Assays to Evaluate Histone Deacetylase 6 Inhibitors.- Immunoprecipitation of HDAC6 and Interacting Proteins.
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