An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives

Abstract 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...

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Autores principales:	Lauren N. Randolph, Xiaoping Bao, Chikai Zhou, Xiaojun Lian
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1d207dfbaee24e02ac21452d78d5023a
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spelling	oai:doaj.org-article:1d207dfbaee24e02ac21452d78d5023a2021-12-02T12:30:16ZAn all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives10.1038/s41598-017-01684-62045-2322https://doaj.org/article/1d207dfbaee24e02ac21452d78d5023a2017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01684-6https://doaj.org/toc/2045-2322Abstract 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.Lauren N. RandolphXiaoping BaoChikai ZhouXiaojun LianNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-8 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lauren N. Randolph Xiaoping Bao Chikai Zhou Xiaojun Lian An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives
description	Abstract 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	article
author	Lauren N. Randolph Xiaoping Bao Chikai Zhou Xiaojun Lian
author_facet	Lauren N. Randolph Xiaoping Bao Chikai Zhou Xiaojun Lian
author_sort	Lauren N. Randolph
title	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_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_sort	all-in-one, tet-on 3g inducible piggybac system for human pluripotent stem cells and derivatives
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/1d207dfbaee24e02ac21452d78d5023a
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An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives

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