A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
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Keywords
amyloid-beta, Alzheimer, cytomegalovirus, fluorescent reporter, high-throughput
Abstract
Robust assays for detecting the effects of elevated concentrations of amyloid-β (Aβ) may facilitate Alzheimer’s disease research. An appropriate assay would be high-throughput and enable identification of drugs and genetic mutations that block the effects of Aβ, potentially leading to treatments for Alzheimer’s disease. We discovered that the commonly used cytomegalovirus (CMV) enhancer/promoter is sensitive to the effects of Aβ. By combining the CMV enhancer/promoter with a fluorescent protein, we created a reporter system that produces changes in intracellular fluorescence in response to Aβ. Using hippocampal neurons, we quantified the ability of a CMV-fluorescent protein recombinant reporter to detect both exogenously applied and overexpressed Aβ. This is the first report of a high-throughput enhancer/promoter-based Aβ detection method. The reporter is able to detect the effects of elevated concentrations of Aβ in a high-throughput fashion, providing a new tool for Alzheimer’s disease research and important knowledge about the commonly used CMV enhancer/promoter.
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