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Related Experiment Videos

Quantitative High-Throughput Screening Using a Coincidence Reporter Biocircuit.

Brittany W Schuck1, Ryan MacArthur1, James Inglese1,2

  • 1National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland.

Current Protocols in Neuroscience
|April 12, 2017
PubMed
Summary
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Reporter-biased artifacts can hinder drug discovery. This study introduces quantitative HTS (qHTS) with coincidence reporters to improve screening accuracy and efficiency for identifying potent small molecules.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • Reporter-biased artifacts negatively impact high-throughput screening (HTS) efficiency and quality.
  • Narrow or single-concentration HTS assays often result in false negatives during primary screening.
  • Developing chemical probes and therapeutics requires high-quality screening methods.

Purpose of the Study:

  • To present protocols for generating a coincidence reporter (CR) biocircuit.
  • To describe the creation of stable cell lines expressing the CR biocircuit.
  • To demonstrate quantitative HTS (qHTS) using CR biocircuits for efficient small molecule identification.

Main Methods:

  • Generation of a coincidence reporter (CR) biocircuit.
  • Establishment of stable cell lines expressing the CR biocircuit.
Keywords:
assay developmentcoincidence reporterquantitative high-throughput screening

Related Experiment Videos

  • Quantitative HTS (qHTS) employing the CR biocircuit.

    • Main Results:

    • The developed protocols enable the generation of CR biocircuits and stable cell lines.
    • qHTS using CR biocircuits effectively identifies high-quality biologically active small molecules.
    • The methods address false negatives and false positives in screening.

    Conclusions:

    • Coincidence reporter technology and qHTS enhance the quality and efficiency of HTS.
    • These protocols facilitate the identification of potent small molecules for therapeutic development.
    • The approach improves the reliability of chemical probe discovery.