High Content Analysis Using Targeted RNAi-Mediated Gene Knockdown: Integrating the Cellomics® ArrayScan® HCS Reader and the Dharmacon siARRAY® Cell Cycle siRNA Library


Small interfering RNAs (siRNAs) are able to effectively down-regulate gene expression in mammalian cells through RNA interference (RNAi). Collections of siRNAs targeting gene families, biological processes, or even entire genomes provide a systems-based approach for identifying the role of particular genes in a cell. RNAi-based gene function analysis is particularly powerful when combined with high content screening (HCS) where multi-parametric assays are used to better define the function of a specific gene target.

In this study, parallel knockdown of 80 genes that are known or predicted to be involved in cell cycle regulation was performed using SMARTpool® reagents from Dharmacon, Inc. Monitoring of four relevant phenotypes (cell cycle phase, nuclear area, cell count, and localization of phospho-ERK) using the ArrayScan® VTI HCS Reader from Cellomics, Inc. identified gene targets involved in cell cycle regulation and cell cycle phase-specific arrest.

The results clearly demonstrate that cell cycle status and phenotypic changes in cell populations can be easily assayed using a highly specific and functional SMARTpool siRNA library followed by data acquisition using high content screening instrumentation and software. In addition, these results show the utility of combining Dharmacon rationally designed SMARTpool reagents and Cellomics' HCS technology for functional mapping of genes in specific disease states or model assays.


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High Content Analysis (460K)