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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doaj-1d207dfbaee24e02ac21452d78d5023a2020-12-07T23:59:28ZengNature Publishing GroupScientific Reports2045-23222017-05-01711810.1038/s41598-017-01684-6An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivativesLauren N. Randolph0Xiaoping Bao1Chikai Zhou2Xiaojun Lian3Department of Biomedical Engineering, Pennsylvania State UniversityDepartment of Chemical and Biological Engineering, University of WisconsinDepartment of Cell and Molecular Biology, Karolinska InstitutetDepartment of Biomedical Engineering, Pennsylvania State UniversityAbstract 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.https://doi.org/10.1038/s41598-017-01684-6 |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lauren N. Randolph Xiaoping Bao Chikai Zhou Xiaojun Lian |
spellingShingle |
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 Scientific Reports |
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 Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2017-05-01 |
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. |
url |
https://doi.org/10.1038/s41598-017-01684-6 |
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