A transposon-mediated system for flexible control of transgene expression in stem and progenitor-derived lineages
Aslam Abbasi Akhtar1, Jessica Molina2, Marina Dutra-Clarke2
1Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA; Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
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.
Related Concept Videos
DNA-only Transposons
The donor site from where the transposon is excised is either degraded or...
Transposons
Conservative Site-specific Recombination and Phase Variation
The recognition sites for Cre recombinase called LoxP...
Transgenic Organisms
Overview of Transposition and Recombination
Reporter Genes


