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Related Experiment Video

Updated: Jan 13, 2026

Sensitive Detection of Proteopathic Seeding Activity with FRET Flow Cytometry
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A high-throughput drug screening assay for anti-tau aggregation using split GFP and flow cytometry.

Omnia M H Ibrahium1, Taiwo A Ademoye2, Jessica S Fortin2

  • 1Purdue Institute of Inflammation, Immunology and Infectious Diseases, Purdue University, West Lafayette, IN, 47907, USA.

Scientific Reports
|October 30, 2025
PubMed
Summary

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We developed a novel cell-based assay using split GFP to monitor tau protein aggregation in real-time. This high-throughput method aids in discovering new therapeutics for neurodegenerative diseases like Alzheimer's disease.

Area of Science:

  • Neuroscience
  • Biochemistry
  • Drug Discovery

Background:

  • Tau protein aggregation is a key feature of neurodegenerative diseases, such as Alzheimer's disease.
  • Developing effective anti-aggregation therapeutics is crucial but challenged by a lack of suitable screening methods.
  • Existing methods often fail to accurately replicate cellular conditions for drug screening.

Purpose of the Study:

  • To develop a high-throughput, cell-based assay for monitoring tau aggregation in living cells.
  • To create a physiologically relevant platform for identifying novel anti-tau aggregation therapeutics.
  • To enable efficient screening of potential drug candidates.

Main Methods:

  • Utilized split GFP technology in suspension-adapted HEK293 cells for real-time tau aggregation monitoring.

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Last Updated: Jan 13, 2026

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  • Co-transfected cells with tau proteins fused to complementary GFP fragments, generating fluorescence upon aggregation.
  • Employed flow cytometry for rapid, quantitative analysis and simultaneous assessment of compound efficacy and cytotoxicity.

    • Main Results:

    • The assay successfully detected tau aggregation in cells without external inducers, attributed to enhanced protein expression.
    • A known urea-based tau aggregation inhibitor demonstrated a dose-dependent reduction in fluorescence, confirming assay sensitivity.
    • The system proved compatible with flow cytometry for high-throughput screening.

    Conclusions:

    • The developed split GFP assay provides a robust and efficient platform for studying tau aggregation.
    • This method significantly advances drug discovery for neurodegenerative diseases by offering a physiologically relevant screening tool.
    • It facilitates the identification of novel therapeutics targeting tau aggregation.