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A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter

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...

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Detalles Bibliográficos
Autores principales: Carrico, Zachary M., Le, Geneva, Malinow, Roberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Journal of Biological Methods 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813829/
https://www.ncbi.nlm.nih.gov/pubmed/29457040
http://dx.doi.org/10.14440/jbm.2017.200
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author Carrico, Zachary M.
Le, Geneva
Malinow, Roberto
author_facet Carrico, Zachary M.
Le, Geneva
Malinow, Roberto
author_sort Carrico, Zachary M.
collection PubMed
description 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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spelling pubmed-58138292018-02-15 A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter Carrico, Zachary M. Le, Geneva Malinow, Roberto J Biol Methods Article 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. Journal of Biological Methods 2017-08-07 /pmc/articles/PMC5813829/ /pubmed/29457040 http://dx.doi.org/10.14440/jbm.2017.200 Text en © 2013-2018 The Journal of Biological Methods, All rights reserved. https://creativecommons.org/licenses/by/3.0/ This work is licensed under a Creative Commons Attribution 3.0 License.
spellingShingle Article
Carrico, Zachary M.
Le, Geneva
Malinow, Roberto
A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
title A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
title_full A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
title_fullStr A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
title_full_unstemmed A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
title_short A fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
title_sort fluorescence assay for detecting amyloid-β using the cytomegalovirus enhancer/promoter
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813829/
https://www.ncbi.nlm.nih.gov/pubmed/29457040
http://dx.doi.org/10.14440/jbm.2017.200
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