Phenotypic Screening, 1st ed. 2018 Methods and Protocols Methods in Molecular Biology Series, Vol. 1787

This volume provides a better understanding of the advancements in phenotypic readouts and improved disease models that generate novel biological insights and recapitulate clinically relevant biology. Chapters focus on phenotypic screening, the use of human cell models, microscopic approaches, assays to measure fat accumulation in C. elegans, the threat response in zebrafish, and protein-protein interactions in plant growth and development. Written in the highly successful Methods in Molecular Biology series format, 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.

Authoritative and practical, Phenotypic Screening: Methods and Protocols aims to make phenotypic screening approaches more accessible to a wide array of researchers throughout the academic and biotech communities.

1.      Nutrient Stress Small-Molecule Screening Platform for Escherichia coli

Sara S. El Zahed, Garima Kumar, Madeline Tong, and Eric D. Brown

2.      Validation and Development of an Escherichia coli Riboflavin Pathway Phenotypic Screen Hit as a Small-molecule Ligand of the Flavin Mononucleotide Riboswitch

Carl J. Balibar, Artjohn Villafania, Christopher M. Barbieri, Nick Murgolo, Terry Roemer, Hao Wang, and John A. Howe

3.      Phenotypic Screening of Small Molecules with Anti-malarial Activity for Three Different Parasitic Life Stages

Nobutaka Kato, Sandra March, Sangeeta N Bhatia, and Matthias Marti

4.      Phenotypic Screening for Inhibitors of a Mutant Thrombopoietin Receptor

Anna Ngo, Ann Koay, Christian Pecquet, Carmen C. Diaconu, David A. Jenkins, Andrew K. Shiau,Stefan N. Constantinescu, and Meng Ling Choong

5.      A Cell-based Assay for Mitotic Spindle Orientation

Elina Glaubke and Holger Bastians

6.      Isolation of Skeletal Muscle Stem Cells for Phenotypic Screens for Modulators of Proliferation

7.      Cell-based Methods to Identify Inducers of Human Pancreatic Beta-cell Proliferation

8.      High-content Imaging Phenotypic Screen for Neurogenesis Using Primary Neural Progenitor Cells

Li Sharon Wu and Jingjun Li

9.      Discovery of Modulators of Adipocyte Physiology Using Fully Functionalized Fragments

Andrea Galmozzi, Christopher G. Parker, Bernard P. Kok, Benjamin F. Cravatt, and Enrique Saez

10.    The Ancient Genetic Networks of Obesity: Whole-animal Automated Screening for Conserved Fat Regulators

Wenfan Ke, Anna Drangowska-Way, Daniel Katz, Karsten Siller, and Eyleen J. O’Rourke

11.    Modulation of Threat Response in Larval Zebrafish

Andrew J. Rennekamp

12.    A PLA-iRoCS Pipeline for the Localization of Protein-Protein Interactions in situ

Taras Pasternak, William Teale, Thorsten Falk, Benedetto Ruperti, and Klaus Palme

13.    High-Dimensional Profiling: The Theta Comparative Cell Scoring Method

Scott J Warchal, John C Dawson, and Neil O Carragher

14.    Accessing the Open PHACTS Discovery Platform with Workflow Tools

15.    Quantitative Prioritization of Tool Compounds for Phenotypic Screening

Yuan Wang and Jeremy L. Jenkins

16.    A Quantitative Systems Pharmacology Approach to Infer Pathways Involved in Complex Disease Phenotypes

Mark E. Schurdak, Fen Pei, Timothy R. Lezon, Diane Carlisle, Robert Friedlander, D. Lansing Taylor, and Andrew M. Stern

17.    Phenotype-based High-content Screening using Fluorescent Chemical Bioprobes: Lipid Droplets and Glucose Uptake Quantification in Live Cells

Young-Hee Shin andSeung Bum Park

18.    Developing High-throughput Electrophysiological Assays for Phenotypic Screening

19.    Testing Susceptibility of Patient Derived Organoid Cultures to Therapies: Pharmacotyping

Richard A. Burkhart, Lindsey A. Baker, and Hervé Tiriac

20.    Techniques to Enable 1536-well Phenotypic Screening

Sinéad Knight, Helen Plant, Lisa McWilliams, and Mark Wigglesworth

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts