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Development of a fluorescent high throughput assay for tau aggregation
Min Liu1, Jake Ni, Kenneth S Kosik
1Laboratory for Drug Discovery in Neurodegeneration, Brigham and Women's Hospital and Harvard Medical School, Cambridge, MA, USA.
Assay and Drug Development Technologies
|January 28, 2005
Summary
A novel high-throughput assay quantifies tau aggregation using fluorescent resonance energy transfer (FRET). This method, utilizing labeled tau and arachidonic acid, can identify potential inhibitors of tau aggregation.
Area of Science:
- Biochemistry
- Neuroscience
- Assay Development
Background:
- Tau aggregation is a key pathological hallmark in neurodegenerative diseases like Alzheimer's.
- Developing efficient methods to study tau aggregation is crucial for understanding disease mechanisms and therapeutic development.
Purpose of the Study:
- To describe a high-throughput assay for measuring tau aggregation.
- To characterize the nature of tau aggregation induced by arachidonic acid.
- To demonstrate the assay's utility in identifying aggregation inhibitors.
Main Methods:
- Utilized full-length recombinant tau labeled with Cy3 or Cy5 dyes.
- Induced tau aggregation with arachidonic acid.
- Measured tau aggregation via fluorescent resonance energy transfer (FRET) in a 384-well format.
- Characterized aggregation using competition and cross-linking experiments.
Main Results:
- Developed and validated a high-throughput FRET assay for tau aggregation.
- Demonstrated that arachidonic acid induces tau aggregation.
- Identified tau dimers and tetramers as the primary species observed via FRET.
- Presented a model for tau aggregation.
Conclusions:
- The FRET assay effectively measures tau aggregation in a high-throughput manner.
- The assay can be employed to screen for compounds that inhibit tau aggregation.
- Understanding tau aggregation is vital for developing treatments for tauopathies.