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A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models

In recent years, there has been an increasing demand for predictive and sensitive in vitro tools for drug discovery. Split complementation assays have the potential to enlarge the arsenal of in vitro tools for compound screening, with most of them relying on well-established reporter gene assays. In...

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Detalles Bibliográficos
Autores principales: Calabretta, Maria Maddalena, Lopreside, Antonia, Montali, Laura, Cevenini, Luca, Roda, Aldo, Michelini, Elisa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7865915/
https://www.ncbi.nlm.nih.gov/pubmed/33572727
http://dx.doi.org/10.3390/s21030893
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author Calabretta, Maria Maddalena
Lopreside, Antonia
Montali, Laura
Cevenini, Luca
Roda, Aldo
Michelini, Elisa
author_facet Calabretta, Maria Maddalena
Lopreside, Antonia
Montali, Laura
Cevenini, Luca
Roda, Aldo
Michelini, Elisa
author_sort Calabretta, Maria Maddalena
collection PubMed
description In recent years, there has been an increasing demand for predictive and sensitive in vitro tools for drug discovery. Split complementation assays have the potential to enlarge the arsenal of in vitro tools for compound screening, with most of them relying on well-established reporter gene assays. In particular, ligand-induced complementation of split luciferases is emerging as a suitable approach for monitoring protein–protein interactions. We hereby report an intracellular nanosensor for the screening of compounds with androgenic activity based on a split NanoLuc reporter. We also confirm the suitability of using 3D spheroids of Human Embryonic Kidney (HEK-293) cells for upgrading the 2D cell-based assay. A limit of detection of 4 pM and a half maximal effective concentration (EC(50)) of 1.7 ± 0.3 nM were obtained for testosterone with HEK293 spheroids. This genetically encoded nanosensor also represents a new tool for real time imaging of the activation state of the androgen receptor, thus being suitable for analysing molecules with androgenic activity, including new drugs or endocrine disrupting molecules.
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spelling pubmed-78659152021-02-07 A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models Calabretta, Maria Maddalena Lopreside, Antonia Montali, Laura Cevenini, Luca Roda, Aldo Michelini, Elisa Sensors (Basel) Article In recent years, there has been an increasing demand for predictive and sensitive in vitro tools for drug discovery. Split complementation assays have the potential to enlarge the arsenal of in vitro tools for compound screening, with most of them relying on well-established reporter gene assays. In particular, ligand-induced complementation of split luciferases is emerging as a suitable approach for monitoring protein–protein interactions. We hereby report an intracellular nanosensor for the screening of compounds with androgenic activity based on a split NanoLuc reporter. We also confirm the suitability of using 3D spheroids of Human Embryonic Kidney (HEK-293) cells for upgrading the 2D cell-based assay. A limit of detection of 4 pM and a half maximal effective concentration (EC(50)) of 1.7 ± 0.3 nM were obtained for testosterone with HEK293 spheroids. This genetically encoded nanosensor also represents a new tool for real time imaging of the activation state of the androgen receptor, thus being suitable for analysing molecules with androgenic activity, including new drugs or endocrine disrupting molecules. MDPI 2021-01-29 /pmc/articles/PMC7865915/ /pubmed/33572727 http://dx.doi.org/10.3390/s21030893 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Calabretta, Maria Maddalena
Lopreside, Antonia
Montali, Laura
Cevenini, Luca
Roda, Aldo
Michelini, Elisa
A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models
title A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models
title_full A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models
title_fullStr A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models
title_full_unstemmed A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models
title_short A Genetically Encoded Bioluminescence Intracellular Nanosensor for Androgen Receptor Activation Monitoring in 3D Cell Models
title_sort genetically encoded bioluminescence intracellular nanosensor for androgen receptor activation monitoring in 3d cell models
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7865915/
https://www.ncbi.nlm.nih.gov/pubmed/33572727
http://dx.doi.org/10.3390/s21030893
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