Autoregulatory tetracycline-regulated system for inducible gene

Details Autoregulatory tetracycline-regulated system for inducible gene Autoregulatory tetracycline-regulated system for inducible gene

1995-06-07

1998-12-22

Fleisher, Mindy

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

4353201, 435325, 435410, 536 234, C12N 1563, C12N 1579, C12N 510, C07H 2104

Patent

active

058517960

ABSTRACT:
A tetracycline-regulated system which provides autoregulatory, inducible gene expression in cultured cells and transgenic animals is described. In the autoregulatory plasmid pTet-tTAk, a modified tTA gene called tTAk was placed under the control of Tetp. Tetracycline prevents tTA from binding to Tetp, preventing expression of both tTA and luciferase. This negative feedback cycle ensures that little or no tTA is produced in the presence of tetracycline, thereby reducing or eliminating possible toxic effects. When tetracycline is removed, however, this strategy predicts that tiny amounts of tTA protein (which may result from the leakiness of the minimal promoter), will bind to Tet-op and stimulate expression of the tTAk gene. A positive feedforward loop is initiated which in turn leads to higher levels of expression of tTA and thus, luciferase. Polynucleotide molecules encoding the autoregulatory system, as well as methods of enhancing or decreasing the expression of desired genes, and kits for carrying out these methods are described.

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