title: High-throughput RNAi screening for the identification of novel targets.
abstract: Gene silencing through RNA interference has provided researchers with an effective way to study gene function. High-throughput RNA interference (HT-RNAi) screening has further permitted researchers to identify functionally relevant mediators of cellular response on a large scale. These screens have greatly expedited the discovery of novel targets and pathway mediators. Here, we describe the methodology for performing HT-RNAi screening of HeLa cells transfected with short interfering RNA (siRNA) libraries in 384-well microplate format. Using this plate format, the HT-RNAi assay can be easily adapted to semi-automated or fully automated platforms. The library siRNA are introduced into the cells through reverse transfection using cationic lipids. HT-RNAi screening for modulators of cell proliferation can be accomplished using a single read out reagent. This type of RNAi screening can be used with most plate-based cellular assays and can be optimized for most cultured cells lines, thus becoming a powerful tool to identify specific gene modulators and targets for drug discovery.
authors: Clinical Translational Research Division, Translational Genomics Research Institute, Scottsdale, AZ, USA.
authors: Meredith C
authors: MC
authors: Henderson
authors: David O
authors: DO
authors: Azorsa
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D058990:Molecular Targeted Therapy; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000032:analysis* / Q000235:genetics*; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering