High-Throughput Rnai Screening: Methods and Protocols

ISBN-13: 9781493963355 / Angielski / Twarda / 2016 / 262 str.

David O. Azorsa; Shilpi Arora

Opis

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.--

Spis treści

1. Establishing an Infrastructure for High-Throughput Short-Interfering RNA Screening
Hongwei Yin, Chris Sereduk, and Nanyun Tang

2. Optimization of Transfection Conditions for siRNA Screening
Justin J. Montoya and David O. Azorsa

3. High Throughput siRNA Screening Using Reverse Transfection
Carina Von Schantz-Fant and Jani Saarela

4. Genome-Wide siRNA Screening Using Forward Transfection: Identification of Modulators of    Membrane Trafficking in Mammalian Cells

Mariana G. Bexiga and Jeremy C. Simpson

5. Pooled shRNA Screening in Mammalian Cells as a Functional Genomic Discovery Platform
Katarzyna Jastrzebski, Bastiaan Evers, and Roderick L. Beijersbergen

6. A Protocol for a High-Throughput Multiplex Cell Viability Assay

Daniel F. Gilbert and Michael Boutros

7. RNAi Screening of Leukemia Cells Using Electroporation
Anupriya Agarwal and Jeffrey W. Tyner

8. Self-Assembled Cell Microarray (SAMcell) for High-Throughput RNAi Screening
Hanshuo Zhang and Juan Li

9. In Vitro Pooled shRNA Screening to Identify Determinants of Radiosensitivity
Alessandro Ceroni, Geoff S. Higgins, and Daniel V. Ebner

10. Three-Dimensional Spheroid Cell Culture Model for Target Identification Utilizing High-Throughput RNAi Screens

Lakesla R. Iles and Geoffrey Bartholomeusz

11. In Vitro High Throughput RNAi Screening to Accelerate the Process of Target Identification and Drug Development
Hongwei Yin and Michelle Kassner

12. High-Throughput, Liquid-Based Genome-Wide RNAi Screening in C. elegans
Linda P. O’Reilly, Ryan R. Knoerdel, Gary A. Silverman, and Stephen C. Pak

13. Design and Methods of Large-Scale RNA Interference Screens in Drosophila
Jia Zhou and Chao Tong

14. Genome-Wide RNAi Screening in C. elegans for Aging and Innate Immunity
Amit Sinha and Robbie Rae

15. RNAi-Assisted Genome Evolution (RAGE) in Saccharomyces cerevisiae
Tong Si and Huimin Zhao

16. RNAi Screening in Spodoptera frugiperda

Subhanita Ghosh, Gatikrushna Singh, Bindiya Sachdev, Ajit Kumar, Pawan Malhotra, Sunil K. Mukherjee, and Raj K. Bhatnagar

17. Morphology and Gene Expression Screening with Morpholinos in Zebrafish Embryos
Li-Chuan Tseng, Chih-Hao Tang and Yun-Jin Jiang

18. Live Cell Microscopy-Based RNAi Screening in the Moss Physcomitrella patens
Tomohiro Miki, Yuki Nakaoka, and Gohta Goshima
19. Data Analysis For High Throughput RNAi Screening
David O. Azorsa, Megan A. Turnidge and Shilpi Arora 

Blurb

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.