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Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription

Type I collagen is a key protein of most connective tissue and its up-regulation is required for wound healing but is also involved in fibrosis. Control of expression of this collagen remains poorly understood apart from Transforming Growth Factor beta (TGF-β1)-mediated induction. To generate a sens...

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Autores principales: Wong, Hui Hui, Seet, Sze Hwee, Bascom, Charles C., Isfort, Robert J., Bard, Frederic
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7665053/
https://www.ncbi.nlm.nih.gov/pubmed/33184327
http://dx.doi.org/10.1038/s41598-020-75683-5
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author Wong, Hui Hui
Seet, Sze Hwee
Bascom, Charles C.
Isfort, Robert J.
Bard, Frederic
author_facet Wong, Hui Hui
Seet, Sze Hwee
Bascom, Charles C.
Isfort, Robert J.
Bard, Frederic
author_sort Wong, Hui Hui
collection PubMed
description Type I collagen is a key protein of most connective tissue and its up-regulation is required for wound healing but is also involved in fibrosis. Control of expression of this collagen remains poorly understood apart from Transforming Growth Factor beta (TGF-β1)-mediated induction. To generate a sensitive, practical, robust, image-based high-throughput-compatible reporter system, we genetically inserted a short-lived fluorescence reporter downstream of the endogenous type I collagen (COL1A1) promoter in skin fibroblasts. Using a variety of controls, we demonstrate that the cell line faithfully reports changes in type I collagen expression with at least threefold enhanced sensitivity compared to endogenous collagen monitoring. We use this assay to test the potency of anti-fibrotic compounds and screen siRNAs for regulators of TGF-β1-induced type I collagen expression. We propose our reporter cell line, Red-COLA1, as a new efficient tool to study type I collagen transcriptional regulation.
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spelling pubmed-76650532020-11-16 Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription Wong, Hui Hui Seet, Sze Hwee Bascom, Charles C. Isfort, Robert J. Bard, Frederic Sci Rep Article Type I collagen is a key protein of most connective tissue and its up-regulation is required for wound healing but is also involved in fibrosis. Control of expression of this collagen remains poorly understood apart from Transforming Growth Factor beta (TGF-β1)-mediated induction. To generate a sensitive, practical, robust, image-based high-throughput-compatible reporter system, we genetically inserted a short-lived fluorescence reporter downstream of the endogenous type I collagen (COL1A1) promoter in skin fibroblasts. Using a variety of controls, we demonstrate that the cell line faithfully reports changes in type I collagen expression with at least threefold enhanced sensitivity compared to endogenous collagen monitoring. We use this assay to test the potency of anti-fibrotic compounds and screen siRNAs for regulators of TGF-β1-induced type I collagen expression. We propose our reporter cell line, Red-COLA1, as a new efficient tool to study type I collagen transcriptional regulation. Nature Publishing Group UK 2020-11-12 /pmc/articles/PMC7665053/ /pubmed/33184327 http://dx.doi.org/10.1038/s41598-020-75683-5 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Wong, Hui Hui
Seet, Sze Hwee
Bascom, Charles C.
Isfort, Robert J.
Bard, Frederic
Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription
title Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription
title_full Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription
title_fullStr Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription
title_full_unstemmed Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription
title_short Red-COLA1: a human fibroblast reporter cell line for type I collagen transcription
title_sort red-cola1: a human fibroblast reporter cell line for type i collagen transcription
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7665053/
https://www.ncbi.nlm.nih.gov/pubmed/33184327
http://dx.doi.org/10.1038/s41598-020-75683-5
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