A 1,536-Well Ultra-High-Throughput siRNA Screen to Identify Regulators of the Wnt/β-Catenin Pathway

• DOI: 10.1089/adt.2009.0262
• Publication Date: 2010-06-26T18:47:51Z
• Authors: Namjin Chung; Shane Marine; Emily A. Smith; Robert Liehr; S. Todd Smith; Louis Locco; Edward Hudak; Anthony Kreamer; Alison Rush; Brian Roberts; Michael B. Major; Randall T. Moon; William Arthur; Michele Cleary; Berta Strulovici; Marc Ferrer
• Keywords: Replicate; High-Throughput Screening

High-throughput siRNA screens are now widely used for identifying novel drug targets and mapping disease pathways. Despite their popularity, there remain challenges related to data variability, primarily due measurement errors, biological variance, uneven transfection efficiency, the efficacy of sequences, or off-target effects, consequent high false discovery rates. Data variability can be reduced if performed in replicate. Running a large-scale screen replicate is difficult, however, because technical automating complicated steps transfection, often with multiplexed assay readouts, controlling environmental humidity during long incubation periods. Small-molecule have greatly benefited past decade from miniaturization high-density plates such that 1,536-well nanoplate screenings routine process, allowing fast, efficient, affordable operations without compromising underlying biology important characteristics. Here, we describe development protocol utilizes instruments commonly found small-molecule throughput screening laboratories. This was then successfully demonstrated triplicate identification regulators Wnt/beta-catenin pathway.