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An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives
Human pluripotent stem cells (hPSCs) offer tremendous promise in tissue engineering and cell-based therapies due to their unique combination of two properties: pluripotency and unlimited proliferative capacity. However, directed differentiation of hPSCs to clinically relevant cell lineages is needed...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5431539/ https://www.ncbi.nlm.nih.gov/pubmed/28484230 http://dx.doi.org/10.1038/s41598-017-01684-6 |
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author | Randolph, Lauren N. Bao, Xiaoping Zhou, Chikai Lian, Xiaojun |
author_facet | Randolph, Lauren N. Bao, Xiaoping Zhou, Chikai Lian, Xiaojun |
author_sort | Randolph, Lauren N. |
collection | PubMed |
description | Human pluripotent stem cells (hPSCs) offer tremendous promise in tissue engineering and cell-based therapies due to their unique combination of two properties: pluripotency and unlimited proliferative capacity. However, directed differentiation of hPSCs to clinically relevant cell lineages is needed to achieve the goal of hPSC-based therapies. This requires a deep understanding of how cell signaling pathways converge on the nucleus to control differentiation and the ability to dissect gene function in a temporal manner. Here, we report the use of the PiggyBac transposon and a Tet-On 3G drug-inducible gene expression system to achieve versatile inducible gene expression in hPSC lines. Our new system, XLone, offers improvement over previous Tet-On systems with significantly reduced background expression and increased sensitivity to doxycycline. Transgene expression in hPSCs is tightly regulated in response to doxycycline treatment. In addition, the PiggyBac elements in our XLone construct provide a rapid and efficient strategy for generating stable transgenic hPSCs. Our inducible gene expression PiggyBac transposon system should facilitate the study of gene function and directed differentiation in human stem cells. |
format | Online Article Text |
id | pubmed-5431539 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-54315392017-05-16 An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives Randolph, Lauren N. Bao, Xiaoping Zhou, Chikai Lian, Xiaojun Sci Rep Article Human pluripotent stem cells (hPSCs) offer tremendous promise in tissue engineering and cell-based therapies due to their unique combination of two properties: pluripotency and unlimited proliferative capacity. However, directed differentiation of hPSCs to clinically relevant cell lineages is needed to achieve the goal of hPSC-based therapies. This requires a deep understanding of how cell signaling pathways converge on the nucleus to control differentiation and the ability to dissect gene function in a temporal manner. Here, we report the use of the PiggyBac transposon and a Tet-On 3G drug-inducible gene expression system to achieve versatile inducible gene expression in hPSC lines. Our new system, XLone, offers improvement over previous Tet-On systems with significantly reduced background expression and increased sensitivity to doxycycline. Transgene expression in hPSCs is tightly regulated in response to doxycycline treatment. In addition, the PiggyBac elements in our XLone construct provide a rapid and efficient strategy for generating stable transgenic hPSCs. Our inducible gene expression PiggyBac transposon system should facilitate the study of gene function and directed differentiation in human stem cells. Nature Publishing Group UK 2017-05-08 /pmc/articles/PMC5431539/ /pubmed/28484230 http://dx.doi.org/10.1038/s41598-017-01684-6 Text en © The Author(s) 2017 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Randolph, Lauren N. Bao, Xiaoping Zhou, Chikai Lian, Xiaojun An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives |
title | An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives |
title_full | An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives |
title_fullStr | An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives |
title_full_unstemmed | An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives |
title_short | An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives |
title_sort | all-in-one, tet-on 3g inducible piggybac system for human pluripotent stem cells and derivatives |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5431539/ https://www.ncbi.nlm.nih.gov/pubmed/28484230 http://dx.doi.org/10.1038/s41598-017-01684-6 |
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