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High Content RNAi Screening Using Printed Libraries下載
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February 04, 2016
Authors: Paul Held, Peter Banks, BioTek Instruments, Inc. Winooski, VT USA; Neil Emans, Persomics USA, Waltham, MA USA
RNA interference is a research tool used to assign function to genes and has proven to be a powerful tool to study gene function by silencing transcription. RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression, typically by causing the destruction of specific mRNA molecules [1-3]. Gene silencing with siRNA is one of a set of functional genomics technologies that enable researchers to identify the precise role a gene plays in a specific biologic process, which is one of the key steps in identifying therapeutic targets with well understood mechanisms of action.
While phenotypic screening is undergoing a resurgence, its use in conjunction with RNAi remains an expensive research strategy, mostly driven by costly assay reagents and the liquid handing and high content imaging instrumentation required to perform screening. Recently, these barriers have been lowered through the development of affordable, high quality automated microscope/imaging systems and the invention of miniaturized RNAi screening technologies that eliminate the requirement for automation infrastructure and reduce cost/data point by orders of magnitude. It is now possible to screen entire RNAi libraries at the bench, eliminating any need for liquid handling robotics.
Persomics technology is fundamentally different than the multi-well microplates that have become the norm in RNAi screening. Persomics plates replace the experimental well with a printed array of spots on an optical glass insert embedded in the base of an SBS standard plate format. Each of these spots is a dried spot of siRNA that also contains all the reagents necessary to silence genes in cells grown over them. This library screening solution requires a limited number of workflow steps for phenotypic screening using a cellular assay and an automated fluorescence imaging system, such as the Cytation. Unlike microplate wells, there is no physical barrier between the individual RNAi experiments, making
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