Selecting the optimal Tet-On system for doxycycline-inducible gene expression in transiently transfected and stably transduced mammalian cells

Biotechnology Journal
Atze T DasB Berkhout

Abstract

The doxycycline (dox)-inducible Tet-On system is widely used to control gene expression in mammalian cells. This system is based on the bacterial Tet operon, which has been modified and improved for its function in eukaryotic cells. To identify the optimal system for different applications, we compared Tet-On variants in frequently used cell types that were either transiently transfected with the relevant plasmids or stably transduced with an "all-in-one" lentiviral vector. The V10 variant performed optimally in the transiently transfected cells and demonstrated no background activity without dox, high dox-induced activity and the highest fold-induction. Because of its very high dox-sensitivity, the V16 system may be preferred if only low intracellular dox concentrations can be reached. V16 performed optimally in the transduced cells and demonstrated the highest activity and dox-sensitivity without background activity. Moreover, V16 demonstrated more robust induction of gene expression after a latency period without dox. This study provides important findings for choosing the optimal Tet-On system for diverse cell culture settings. V10 is the best system for most applications in which the DNA is episomally present in cells, whe...Continue Reading

References

Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M Gossen, H Bujard
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May 17, 2001·Proceedings of the National Academy of Sciences of the United States of America·G MarzioB Berkhout
Feb 6, 2004·The Journal of Biological Chemistry·Atze T DasBen Berkhout
Jun 23, 2012·Nature·Masaharu KinoshitaTadashi Isa
Oct 17, 2013·Frontiers in Neural Circuits·Thongchai SooksawateTadashi Isa

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Citations

Jun 17, 2016·Scientific Reports·Yuechuan ZhangDeqiang Wang
Jul 11, 2018·Biotechnology Journal·Alan CostelloMartin Clynes
Aug 4, 2021·Applied Microbiology and Biotechnology·Aniel Moya-TorresJörg Stetefeld
Aug 11, 2021·Development, Growth & Differentiation·Osamu HosoyaMary Miyaji

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