Tetracycline-Regulatable Gene Transcription Switch

Brigham and Women's Hospital
posted on 08/12/2011

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Technology: Dr. Yao has developed a powerful repression-based transcription switch that exploits the bacterial operon controlling tetracycline resistance to turn expression of recombinant DNA on and off in mammalian cells. The bacterial operon comprises promoter and operator elements that control expression of two proteins: the tetR protein and the operon repressor protein. In the absence of tetracycline, the repressor protein binds to the operator DNA sequence and prevents transcription of both genes. In the presence of tetracycline, the drug binds the repressor protein and causes it to detach from the operator sequence. As a result, RNA polymerase is able to bind to the promoter sequence and the tetR gene and repressor are transcribed. Applications: - Therapeutic protein and antibody production - Therapeutic vaccine production - Drug screening Advantages: - Tightly controlled inducible gene expression. - Increased therapeutic protein production in mammalian cells. - Prevention of endogenous and latent viral replication when the transcription switch is integrated into vaccine constructs. - Allows for the stable expression of target gene products that are toxic to cells, by permitting expansion of the cells to high density prior to expressing the target gene product.