High-Throughput RNAi Screening

High-throughput RNAi screening remains one of the most widely used technologies to perform target identification and validation studies in an unbiased manner. These assays are equally important for research and development across academic, biotech, and pharmaceutical industries. The success of these screening efforts is dependent on robust methodologies to perform these screens. In High-Throughput RNAi Screening: Methods and Protocols, expert researchers in the field share protocols and methods for performing high-throughput RNAi (HT-RNAi) screens. These include the use of various RNAi platforms and delivery methods in mammalian and non-mammalian systems, whole organism and cell models, and various applications, such as drug sensitizer identification. Finally, the book examines the latest advancements in the fields of assay development, library screening, data analysis, and hit selection. 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.

Cutting-edge and thorough, High-Throughput RNAi Screening: Methods and Protocols provides a comprehensive source of protocols and other necessary information to make robust and successful assays possible for all who wish to apply HT-RNAi in their research.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Establishing an Infrastructure for High-Throughput Short-Interfering RNA Screening.- Optimization of Transfection Conditions for siRNA Screening.- High Throughput siRNA Screening Using Reverse Transfection.- Genome-Wide siRNA Screening Using Forward Transfection: Identification of Modulators of Membrane Trafficking in Mammalian Cells.- Pooled shRNA Screening in Mammalian Cells as a Functional Genomic Discovery Platform.- A Protocol for a High-Throughput Multiplex Cell Viability Assay.- RNAi Screening of Leukemia Cells Using Electroporation.- Self-Assembled Cell Microarray (SAMcell) for High-Throughput RNAi Screening.- In Vitro Pooled shRNA Screening to Identify Determinants of Radiosensitivity.- Three-Dimensional Spheroid Cell Culture Model for Target Identification Utilizing High-Throughput RNAi Screens.- In Vitro High Throughput RNAi Screening to Accelerate the Process of Target Identification and Drug Development.- High-Throughput, Liquid-Based Genome-Wide RNAi Screening in C. elegans.- Design and Methods of Large-Scale RNA Interference Screens in Drosophila.- Genome-Wide RNAi Screening in C. elegans for Aging and Innate Immunity.- RNAi-Assisted Genome Evolution (RAGE) in Saccharomyces cerevisiae.- RNAi Screening in Spodoptera frugiperda.- Morphology and Gene Expression Screening with Morpholinos in Zebrafish Embryos.- Live Cell Microscopy-Based RNAi Screening in the Moss Physcomitrella patens.- Data Analysis For High Throughput RNAi Screening.