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 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.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Randolph LN,Bao X,Zhou C,Lian X

doi

10.1038/s41598-017-01684-6

subject

Has Abstract

pub_date

2017-05-08 00:00:00

pages

1549

issue

1

issn

2045-2322

pii

10.1038/s41598-017-01684-6

journal_volume

7

pub_type

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