Cargando…
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...
Autores principales: | , , , , |
---|---|
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 |
_version_ | 1783609949131112448 |
---|---|
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. |
format | Online Article Text |
id | pubmed-7665053 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT wonghuihui redcola1ahumanfibroblastreportercelllinefortypeicollagentranscription AT seetszehwee redcola1ahumanfibroblastreportercelllinefortypeicollagentranscription AT bascomcharlesc redcola1ahumanfibroblastreportercelllinefortypeicollagentranscription AT isfortrobertj redcola1ahumanfibroblastreportercelllinefortypeicollagentranscription AT bardfrederic redcola1ahumanfibroblastreportercelllinefortypeicollagentranscription |