Introduction

Tetracycline (TET) technology allows precise, reversible, and efficient spatiotemporal control of gene expression. This “on demand” gene induction mimics disease onset and disease progression. When coupled with Cre recombinase, TET technology allows one to selectively shut down target gene expression.  

There are two most commonly used inducible expression systems of research of eukaryote cell biology: Tet-Off and Tet-On. The Tet-Off system makes use of the tetracycline transactivator (tTA) protein, which is created by fusing TetR (tetracycline repressor), with the activation domain of VP16. The resulting tTA protein is able to bind to DNA at specific TetO operator sequences. In most Tet-off systems, several repeats of such TetO sequences are placed upstream of a minimal promoter such as the CMV promoter. In a Tet-Off system, expression of TRE-controlled genes can be repressed by tetracycline and its derivatives. They bind tTA and render it incapable of binding to TRE sequences, thereby preventing transactivation of TRE-controlled genes.

The advantage of the Tet system over other conditional gene expression systems (e.g. Cre, FRT, and ER) is that in other systems, the activation or knockout of the gene is irreversible once recombination is accomplished, but in Tet system, it’s reversible.

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