Identification of Therapeutic Small-Molecule Leads in Cultured Cells Using Multiplexed Pathway Reporter Readouts
The rapid expansion of molecular screening libraries in size and complexity in the last decade has outpaced the discovery rate of cost-effective strategies to single out reagents with sought-after cellular activities. In addition to representing high-priority therapeutic targets, intensely studied cell signaling systems encapsulate robust reference points for mapping novel chemical activities given our deep understanding of the molecular mechanisms that support their activity. In this chapter, we describe strategies for using transcriptional reporters of several well-interrogated signal transduction pathways coupled with high-throughput biochemical assays to fingerprint novel compounds for drug target identification agendas.
Key wordsSmall-molecule screening RNAi Luciferase assay Wnt TP53 Kras Dot blotting
This work was supported by the NIH (R01 CA168761), CPRIT (RP130212), and the Welch Foundation (I-1665). This work was also supported by the fund from Japan Society for the Promotion of Science (JSPS) for the “Institutional Program for Young Researcher Overseas Visits.”
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