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HDAC/HAT Function Assessment and Inhibitor Development Methods and Protocols

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Lingua: Inglese


Pubblicazione: 10/2022
Edizione: 2nd ed. 2023


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.


Part I: In Vivo and 3R Models to Assess HDAC Functions and Effects of HDACi


1. Analyzing Lymphoma Development and Progression Using HDACi in Mouse Models

            Eva-Maria Piskor, René Winkler, and Christian Kosan


2. Colony Formation Assay to Test the Impact of HDACi on Leukemic Cells

            Miriam Pons and Mandy Beyer


3. Defined Human Leukemic CD34+ Liquid Cultures to Study HDAC/Transcriptional Repressor Complexes

            Roland Windisch, Sophie Kreissig, and Christian Wichmann


4. Development of a Cellular Model Mimicking Specific HDAC Inhibitors

            Lena Hess, Verena Moos, and Christian Seiser


5. Evaluation of Anti-Tumor and On-Target Activity of HDAC Inhibitors with the Zebrafish Embryo Xenograft Model

            Charlotte Gatzweiler, Johannes Ridinger, Simay Ayhan, Sara Najafi, Heike Peterziel, Olaf Witt, and Ina Oehme


6. Human Platelet Lysate as Valid Cell Growth Additive to Assess Protein Acetylation

            Miriam Pons, Roland Conradi, Walburgis Brenner, and Oliver H. Krämer


7. Investigating Physiopathological Roles for Sirtuins in a Mouse Model

            Shimin Sun, Xiaojiao Xia, Ming Wang, and Baohua Liu


8. Patient-Derived Organoids for In Vivo Validation of In Vitro Data

            Said Kdimati, Florian Bürtin, Michael Linnebacher, and Christina Susanne Mullins


Part II: Novel HDAC Inhibitors and Improved Formulations for HDAC Inhibitors


9. Analysis of HDACi-Coupled Nanoparticles: Opportunities and Challenges

            Marie Kühne, Susanne Hofmann, Henry Lindemann, Zoltán Cseresnyés, Andreas Dzierza, Daniel Schröder, Maren Godmann, Andreas Koschella, Christian Eggeling, Dagmar Fischer, Marc Thilo Figge, Thomas Heinze, and Thorsten Heinzel


10. Development of Pyrazine-Anilinobenzamides as Histone Deacetylase HDAC1-3 Selective Inhibitors and Biological Testing Against Pancreas Cancer Cell Lines

            Mohamed Abdelsalam, Hany S. Ibrahim, Lukas Krauss, Matthes Zessin, Anita Vecchio, Sieglinde Hastreiter, Mike Schutkowski, Günter Schneider, and Wolfgang Sippl


11. Evaluation of Small Molecule HDAC Inhibitors through In Vitro and In Cellulo Approaches

            Nabanita Nawar, Harsimran Kaur Garcha, Abootaleb Sedighi, Elvin D. de Araujo, and Patrick T. Gunning


12. Synthesis, Biochemical, and Cellular Evaluation of HDAC6 Targeting Proteolysis Targeting Chimeras

            Salma Darwish, Tino Heimburg, Johannes Ridinger, Daniel Herp, Matthias Schmidt, Christophe Romier, Manfred Jung, Ina Oehme, and Wolfgang Sippl


13. HDACi Delivery Systems Based on Cellulose Valproate Nanoparticles

            Henry Lindemann, Marie Kühne, Andreas Koschella, Maren Godmann, Thorsten Heinzel, and Thomas Heinze


14. Synthesis and Characterization of Reversible Covalent HDAC4 Inhibitors

            Anton Frühauf, Benjamin Wolff, Markus Schweipert, and Franz-Josef Meyer-Almes


Part III: Innovative Techniques to Determine the Impact of Acetylation on Chromatin, DNA Damage, and Cell Stress Responses


15. Analysis of Cell Cycle and DNA Compaction Dependent Subnuclear Distribution of Histone Marks

            Sunil Kumar Pradhan and M. Cristina Cardoso


16. Analyzing the Effects of HDAC Inhibitors on DNA Damage and Associated Cytotoxicity in Primary Hepatocytes

            Max J. Carlsson and Jörg Fahrer


17. Assessment of HDAC Inhibitor-Induced Endoplasmic Reticulum (ER) Stress

            Melisa Halilovic, Lisa Marx-Blümel, Christian Marx, Katrin Buder, James F. Beck, and Jürgen Sonnemann


18. Assessment of Mitochondrial Dysfunctions after Sirtuin Inhibition

            Christian Marx, Lisa Marx-Blümel, Jürgen Sonnemann, and Zhao-Qi Wang


19. Assessing the Effect of Histone Deacetylase Inhibitors on DNA Double-Strand Break Repair by Non-Homologous End Joining

            Oliver H. Krämer, Tamara Diehl, and Wynand P. Roos


20. Assessing SIRT7 Activity In Vivo and In Vitro in Response to DNA Damage

            Arif Ali Chishti, Zhenhai Li, Baohua Liu, and Wei-Guo Zhu


21. Methods to Evaluate the Effects of HAT/KAT Inhibition on SIAH2-Driven Reactive Oxygen Species Generation in Helicobacter pylori-Infected Gastric Epithelial Cells

            Pragyesh Dixit, Shrikant Babanrao Kokate, Suvasmita Rath, Lopamudra Das, Debashish Chakraborty, and Asima Bhattacharyya


22. Monitoring Changes in Intracellular Reactive Oxygen Species Levels in Response to Histone Deacetylase Inhibitors

            Al-Hassan M. Mustafa, Ramy Ashry, and Oliver H. Krämer


23. Single-Cell Analysis of Histone Acetylation Dynamics at Replication Forks Using PLA and SIRF

            Seo Yun Lee, Jae Jin Kim, and Kyle M. Miller


24. Testing the Effect of Histone Acetyltransferases on Local Chromatin Compaction

            Maximilian Pfisterer and M. Lienhard Schmitz


Part IV: Modern Assays to Evaluate HDAC Functions and HDACi, Including CRISPR-Cas9 Techniques


25. A Genome-Wide CRISPR-Cas9 Loss-of-Function Screening to Identify Host Restriction Factors Modulating Oncolytic Virotherapy

            Michela Muscolini, John Hiscott, and Evelyne Tassone


26. Cloning Strategy for HDAC1/HDAC2 Hybrid Protein Expression in Mammalian Cells

            Désirée Gül, Sandra Olf, Jan Hagemann, Roland H. Stauber, and Oliver H. Krämer


27. Continuous Histone Deacylase Activity Assays

            Matthes Zessin, Marat Meleshin, Wolfgang Sippl, and Mike Schutkowski


28. CRISPR Activation/Interference Screen to Identify Genetic Netwo

Altre Informazioni



Condizione: Nuovo
Collana: Methods in Molecular Biology
Dimensioni: 254 x 178 mm Ø 1186 gr
Formato: Copertina rigida
Illustration Notes:XVII, 511 p. 137 illus., 98 illus. in color.
Pagine Arabe: 511
Pagine Romane: xvii

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