Full 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 fieldsthat need a quick overview of these state-of-the-art technologies.
Contents
Using Functional Genetics in Haploid Cells for Drug Target Identification.- Unbiased Forward Genetic Screening with Chemical Mutagenesis to Uncover Drug-Target Interactions.- Integration of RNAi and Small Molecule Screens to Identify Targets for Drug Development.- CellProfiler and KNIME: Open Source Tools for High Content Screening.- In Silico Target Druggability Assessment: From Structural to Systemic Approaches.- In Silico Drug-Target Profiling.- Locus-Specific Knock-In of a Degradable Tag for Target Validation Studies.- Expression of Human VH Single Domains as Fc-Fusions in Mammalian Cells.- The NeuroSphere Assay (NSA) Applied to Neural Stem Cells (NSCs) and Cancer Stem Cells (CSCs).- 3D-3 Tumor Models in Drug Discovery for Analysis of Immune Cell Infiltration.- Establishment and Analysis of a 3D Co-Culture Spheroid Model of Pancreatic Adenocarcinoma for Application in Drug Discovery.- In Vivo Pharmacology Models for Cancer Target Research.- Use of CRISPR/Cas9 for the Modification of the Mouse Genome.- Assessment of Gene Function of Mouse Innate Lymphoid Cells for In Vivo Analysis Using Retroviral Transduction.- Creation of PDX-Bearing Humanized Mice to Study Immuno-Oncology.- Immunophenotyping of Tissue Samples Using Multicolour Flow Cytometry.- Combined MicroRNA In Situ Hybridization and Immunohistochemical Detection of Protein Markers.- Tangential Flow Filtration with or without Subsequent Bind Elute Size Exclusion Chromatography for Purification of Extracellular Vesicles.- Chromogenic Tissue-Based Methods for Detection of Gene Amplifications (or Rearrangements) Combined with Protein Overexpression in Clinical Samples.
