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A transposon-mediated system for flexible control of transgene expression in stem and progenitor-derived lineages.

Abstract：

:Precise methods for transgene regulation are important to study signaling pathways and cell lineages in biological systems where gene function is often recycled within and across lineages. We engineered a genetic toolset for flexible transgene regulation in these diverse cellular contexts. Specifically, we created an optimized piggyBac transposon-based system, allowing for the facile generation of stably transduced cell lineages in vivo and in vitro. The system, termed pB-Tet-GOI (piggyBac-transposable tetracycline transactivator-mediated flexible expression of a genetic element of interest), incorporates the latest generation of tetracycline (Tet) transactivator and reverse Tet transactivator variants--along with engineered mutants--in order to provide regulated transgene expression upon addition or removal of doxycycline (dox). Altogether, the flexibility of the system allows for dox-induced, dox-suppressed, dox-resistant (i.e., constitutive), and dox-induced/constitutive regulation of transgenes. This versatile strategy provides reversible temporal regulation of transgenes with robust inducibility and minimal leakiness.

journal_name

Stem Cell Reports

journal_title

Stem cell reports

authors

Akhtar AA,Molina J,Dutra-Clarke M,Kim GB,Levy R,Schreiber-Stainthorp W,Danielpour M,Breunig JJ

doi

10.1016/j.stemcr.2015.01.013

subject

Has Abstract

pub_date

2015-03-10 00:00:00

pages

323-31

issue

3

issn

2213-6711

pii

S2213-6711(15)00034-X

journal_volume

4

pub_type

杂志文章

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