The reverse tetracycline-controlled transactivator rtTA2s-S2 is toxic in mouse embryonic stem cells.
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 rtTA2s-S2. We show that the rtTA2s-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 rtTA2s-S2 and neomycin phosphotransferase, we found that embryonic stem cells expressing rtTA2s-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.
Bryja
Vitezslav
V
Center for Cell Therapy and Tissue Repair, Charles University, Department of Molecular Embryology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
Pachernik
Jiri
J
Kubala
Lukas
L
Hampl
Ales
A
Dvorak
Petr
P
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010957:Plasmids / Q000235:genetics / Q000502:physiology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*; D013752:Tetracycline / Q000494:pharmacology*; D015534:Trans-Activators / Q000235:genetics / Q000494:pharmacology*; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D015534:Trans-Activators; D008156:Luciferases; D001616:beta-Galactosidase; D013752:Tetracycline