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Related Experiment Videos

A reporter system for reverse transfection cell arrays.

Brian L Webb1, Begoña Díaz, G Steven Martin

  • 1Science and Technology Division, Corning Incorporated, Corning, NY 14832, USA. webbbl@corning.com

Journal of Biomolecular Screening
|January 9, 2004
PubMed
Summary

Reverse transfection microarrays enable high-throughput gene function screening. A novel reporter system using green fluorescent protein (GFP) monitors mitogen-activated protein (MAP) kinase signaling, accelerating drug discovery.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Pharmacology

Background:

  • Genomic advancements necessitate high-throughput gene discovery methods.
  • Conventional methods struggle to keep pace with rapid gene sequencing.
  • Reverse transfection offers a high-throughput solution for in vivo gene function studies.

Purpose of the Study:

  • To develop a reporter system for monitoring gene function in reverse transfection microarrays.
  • To enable parallel screening of hundreds to thousands of genes.
  • To overcome limitations in posttransfection processing for gene expression analysis.

Main Methods:

  • Developed a serum response element (SRE) reporter linked to green fluorescent protein (GFP).
  • Cotransfected the SRE-GFP reporter with target genes on reverse transfection arrays.

Related Experiment Videos

  • Utilized the system to monitor mitogen-activated protein (MAP) kinase signaling.
  • Applied MEK inhibitor U0126 to test reporter system sensitivity.

    • Main Results:

    • The SRE-GFP reporter system successfully monitored MAP kinase signaling.
    • The system detected inhibition of upstream MAP kinase signaling proteins by U0126.
    • Demonstrated parallel monitoring of multiple signaling proteins in a multiwell format.

    Conclusions:

    • Reverse transfection reporter arrays are valuable for high-throughput screening.
    • This method accelerates the analysis of gene function and signaling pathways.
    • Facilitates rapid drug discovery and development in the pharmaceutical industry.