Reprod. Nutr. Dev.
Volume 43, Number 6, November-December 2003
|Page(s)||477 - 486|
The reverse tetracycline-controlled transactivator rtTA2 -S2 is toxic in mouse embryonic stem cellsVitezslav Bryjaa, b, Jiri Pachernika, c, Lukas Kubalad, Ales Hampla, b, c and Petr Dvoraka, b, c
a Center for Cell Therapy and Tissue Repair, Charles University, V uvalu 84, 150 06, Prague, Czech Republic
b Department of Molecular Embryology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20, Prague, Czech Republic
c Laboratory of Molecular Embryology, Mendel University Brno, Zemedelska 1, 613 00, Brno, Czech Republic
d Laboratory of Free Radical Pathophysiology, Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, 612 65, Brno, Czech Republic
Received 10 February 2003; accepted 10 September 2003
Abstract - The efficient and reversible control of transgene expression is a powerful tool for the correct manipulation of embryonic stem cells in both cell therapy and transgenesis. The aim of this work was to investigate the possibilities of recently developed reverse tetracycline-controlled transactivator rtTA2 -S2. We show that the rtTA2 -S2 is useful for transient inducible expression of genes in embryonic stem cells. However, we found that it was not possible to establish mouse embryonic stem cell lines stably expressing this transactivator. Using the viral IRES sequence which couples the expression of rtTA2 -S2 and neomycin phosphotransferase, we found that embryonic stem cells expressing rtTA2 -S2 are not capable of growing in the presence of G418. Our results indicate that this transactivator is toxic to ES cells and raise the need for the development of other strategies for stable and inducible expression of genes in ES cells.
Key words: TetOn system / reverse tetracycline-controlled transactivator / embryonic stem cell / toxicity
Corresponding author: Petr Dvorak firstname.lastname@example.org
© INRA, EDP Sciences 2003