Regulation of exogenous gene expression in fish cells: An evaluation of different versions of the tetracycline-regulated system

The exogenous control of foreign gene expression is relevant to both basic research and biotechnological applications. In fish, the number of isolated genes has become larger in the last few years; however an efficient system for controlling gene expression is not yet available. The tetracycline-regulated system has proved to be efficient and it is widely used in mammals, but it has never been tested in fish. This work includes the establishment of the tetracycline-regulated system for use in fish cells, and the determination of the optimal conditions to achieve a tight exogenous expression regulation. We have compared the tet-off and tet-on systems and the performance of the transactivators under the control of promoters with different origin and strength. The results show that the tet-off is more efficient than the tet-on system for use in fish cells. The hCMV promoter/enhancer proved to be more efficient than the carp β-actin promoter to drive the expression of the transactivator, since the use of the carp β-actin promoter resulted in a high intra-clonal variability when stably expressed. An auto-regulated system approach proved useful only when transiently expressed.This work was supported by the EC grant FAIR CT98-3482 and the MCYT grant MAR-1999-1647 CE. I.M. was supported by a fellowship from Diputación de Castellón