Cell line for screening demethylating agents using an endogenous epigenetically silenced reporter
The invention provides a high-throughput screening system for agents that influence DNA methylation.
Using the Zinc Finger Nuclease (ZFN) technique, EGFP and G418 resistance genes were stably integrated in the genome of a human cell line under the control of an endogenous promoter. Since this promoter is methylated (epigenetically silenced), the reporter genes are not expressed. After the addition of demethylating agents GFP or G418 can be used as a readout in a screening assay for epigenetic reactivation. The system has a z-value of 0.75 using G418 as readout.
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