Main 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.

Contents:

Part I: Identification of Novel Targets Using High Throughput Screening Assays

1. Using Functional Genetics in Haploid Cells for Drug Target Identification

Jennifer C. Volz, Nicole Schuller, and Ulrich Elling

2. Unbiased Forward Genetic Screening with Chemical Mutagenesis to Uncover Drug-Target Interactions

Moritz Horn, Franziska Metge, and Martin S. Denzel

3. Integration of RNAi and Small Molecule Screens to Identify Targets for Drug Development

Konstantinos Drosopoulos and Spiros Linardopoulos

4. CellProfiler and KNIME: Open Source Tools for High Content Screening

Martin Stoeter, Antje Janosch, Rico Barsacchi, and Marc Bickle

Part II: Drug Target Profiling and Validation

5. In Silico Target Druggability Assessment: From Structural to Systemic Approaches

Jean-Yves Trosset and Christian Cave

6. In Silico Drug-Target Profiling

Jean-Yves Trosset and Christian Cave

7. Locus-Specific Knock-In of a Degradable Tag for Target Validation Studies

Matthias Brand and Georg E. Winter

8. Expression of Human VH Single Domains as Fc-Fusions in Mammalian Cells

Mahmoud Abdelatti, Peter Schofield, and Daniel Christ

Part III: Three Dimensional Cell Culture Techniques Mimicking the Tumor Microenvironment In Vitro

9. The NeuroSphere Assay (NSA) Applied to Neural Stem Cells (NSCs) and Cancer Stem Cells (CSCs)

Rossella Galli

10. 3D-3 Tumor Models in Drug Discovery for Analysis of Immune Cell Infiltration

Annika Osswald, Viola Hedrich, and Wolfgang Sommergruber

11. Establishment and Analysis of a 3D Co-Culture Spheroid Model of Pancreatic Adenocarcinoma for Application in Drug Discovery

Julia C. Meier-Hubberten and Michael P. Sanderson

Part IV: Animal Models for the Study of Gene Function In Vivo

12. In Vivo Pharmacology Models for Cancer Target Research

Dawei Chen, Xiaoyu An, Xuesong Ouyang, Jie Cai, Demin Zhou, and Qi-Xiang Li

13. Use of CRISPR/Cas9 for the Modification of the Mouse Genome

Alexander Klimke, Steffen Guttler, Petric Kuballa, Simone Janzen, Sonja Ortmann, and Adriano Flora

14. Assessment of Gene Function of Mouse Innate Lymphoid Cells for In Vivo Analysis Using Retroviral Transduction

Cyril Seillet and Gabrielle T. Belz

15. Creation of PDX-Bearing Humanized Mice to Study Immuno-Oncology

Li-Chin Yao, Ken-Edwin Aryee, Mingshan Cheng, Pali Kaur, James G. Keck, and Michael A. Brehm

16. Immunophenotyping of Tissue Samples Using Multicolour Flow Cytometry

Martina M. Sykora and Markus Reschke

Part V: Translational Methods to Validate Biomarkers

17. Combined MicroRNA In Situ Hybridization and Immunohistochemical Detection of Protein Markers

Boye Schnack Nielsen and Kim Holmstrom

18. 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, and Samir El Andaloussi

19. Chromogenic Tissue-Based Methods for Detection of Gene Amplifications (or Rearrangements) Combined with Protein Overexpression in Clinical Samples

Hiroaki Nitta and Brian Kelly