The tetracycline-controlled Tet-Off and Tet-On gene expression systems are used to regulate
the activity of genes in eukaryotic cells in diverse settings, varying from basic biological research
to biotechnology and gene therapy applications. These systems are based on regulatory elements that
control the activity of the tetracycline-resistance operon in bacteria. The Tet-Off system allows silencing
of gene expression by administration of tetracycline (Tc) or tetracycline-derivatives like doxycycline
(dox), whereas the Tet-On system allows activation of gene expression by dox. Since the initial design and construction
of the original Tet-system, these bacterium-derived systems have been significantly improved for their function in
eukaryotic cells. We here review how a dox-controlled HIV-1 variant was designed and used to greatly improve the activity
and dox-sensitivity of the rtTA transcriptional activator component of the Tet-On system. These optimized rtTA variants
require less dox for activation, which will reduce side effects and allow gene control in tissues where a relatively low
dox level can be reached, such as the brain.
Keywords: Tet-On system, Tet-Off system, Transcription regulation, Doxycycline, Gene expression, rtTA, tTA, TetR.
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