Identification of Therapeutic Small-Molecule Leads in Cultured Cells Using Multiplexed Pathway Reporter Readouts

  • Ozlem Kulak
  • Kiyoshi Yamaguchi
  • Lawrence Lum
Part of the Methods in Molecular Biology book series (MIMB, volume 1263)


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 words

Small-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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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Cell BiologyUniversity of Texas Southwestern Medical CenterDallasUSA
  2. 2.Division of Clinical Genome Research, Advanced Clinical Research Center, Institute of Medical ScienceUniversity of TokyoTokyoJapan

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