期刊
BIOTECHNOLOGY JOURNAL
卷 11, 期 1, 页码 71-79出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201500236
关键词
Doxycycline; Gene expression; rtTA; Tet-On system; Transcription regulation
资金
- Technology Foundation STW (applied science division of the Netherlands Organisation for Scientific Research NWO)
- Technology Foundation STW (the technology program of the Ministry of Economic Affairs, Utrecht, the Netherlands)
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, whereas V16 may be optimal when the Tet-On components are stably integrated in the cellular genome.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据