Target Identification and Validation in Drug Discovery: Methods and Protocols

Description

This second edition book explores breakthrough technologies in the field of drug target identification and validation. The volume emphasizes particularly revolutionary technologies, such as CRISPR-related screening, “big data,” and in silico approaches, as well as in vivo applications of CRISPR and best uses of animal models in drug development. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Fully updated and authoritative, Target Identification and Validation in Drug Discovery: Methods and Protocols, Second Edition is an ideal guide for molecular and cellular biologists, pharmacologists, pathologists, bioinformaticians, clinical researchers, or investigators, as well as experts in other fields that need a quick overview of these state-of-the-art technologies. Front Matter ....Pages i-xiii Front Matter ....Pages 1-1 Using Functional Genetics in Haploid Cells for Drug Target Identification (Jennifer C. Volz, Nicole Schuller, Ulrich Elling)....Pages 3-21 Unbiased Forward Genetic Screening with Chemical Mutagenesis to Uncover Drug–Target Interactions (Moritz Horn, Franziska Metge, Martin S. Denzel)....Pages 23-31 Integration of RNAi and Small Molecule Screens to Identify Targets for Drug Development (Konstantinos Drosopoulos, Spiros Linardopoulos)....Pages 33-42 CellProfiler and KNIME: Open-Source Tools for High-Content Screening (Martin Stöter, Antje Janosch, Rico Barsacchi, Marc Bickle)....Pages 43-60 Front Matter ....Pages 61-61 In Silico Target Druggability Assessment: From Structural to Systemic Approaches (Jean-Yves Trosset, Christian Cavé)....Pages 63-88 In Silico Drug–Target Profiling (Jean-Yves Trosset, Christian Cavé)....Pages 89-103 Locus-Specific Knock-In of a Degradable Tag for Target Validation Studies (Matthias Brand, Georg E. Winter)....Pages 105-119 Expression of Human VH Single Domains as Fc Fusions in Mammalian Cells (Mahmoud Abdelatti, Peter Schofield, Daniel Christ)....Pages 121-136 Front Matter ....Pages 137-137 The Neurosphere Assay (NSA) Applied to Neural Stem Cells (NSCs) and Cancer Stem Cells (CSCs) (Rossella Galli)....Pages 139-149 3D-3 Tumor Models in Drug Discovery for Analysis of Immune Cell Infiltration (Annika Osswald, Viola Hedrich, Wolfgang Sommergruber)....Pages 151-162 Establishment and Analysis of a 3D Co-Culture Spheroid Model of Pancreatic Adenocarcinoma for Application in Drug Discovery (Julia C. Meier-Hubberten, Michael P. Sanderson)....Pages 163-179 Front Matter ....Pages 181-181 In Vivo Pharmacology Models for Cancer Target Research (Dawei Chen, Xiaoyu An, Xuesong Ouyang, Jie Cai, Demin Zhou, Qi-Xiang Li)....Pages 183-211 Use of CRISPR/Cas9 for the Modification of the Mouse Genome (Alexander Klimke, Steffen Güttler, Petric Kuballa, Simone Janzen, Sonja Ortmann, Adriano Flora)....Pages 213-230 Assessment of Gene Function of Mouse Innate Lymphoid Cells for In Vivo Analysis Using Retroviral Transduction (Cyril Seillet, Gabrielle T. Belz)....Pages 231-240 Creation of PDX-Bearing Humanized Mice to Study Immuno-oncology (Li-Chin Yao, Ken-Edwin Aryee, Mingshan Cheng, Pali Kaur, James G. Keck, Michael A. Brehm)....Pages 241-252 Immunophenotyping of Tissue Samples Using Multicolor Flow Cytometry (Martina M. Sykora, Markus Reschke)....Pages 253-268 Front Matter ....Pages 269-269 Combined MicroRNA In Situ Hybridization and Immunohistochemical Detection of Protein Markers (Boye Schnack Nielsen, Kim Holmstrøm)....Pages 271-286 Tangential Flow Filtration with or Without Subsequent Bind-Elute Size Exclusion Chromatography for Purification of Extracellular Vesicles (Joel Z. Nordin, R. Beklem Bostancioglu, Giulia Corso, Samir EL Andaloussi)....Pages 287-299 Chromogenic Tissue-Based Methods for Detection of Gene Amplifications (or Rearrangements) Combined with Protein Overexpression in Clinical Samples (Hiroaki Nitta, Brian Kelly)....Pages 301-314 Back Matter ....Pages 315-318