The reverse tetracycline-controlled transactivator rtTA2(S)-S2 is toxic in mouse embryonic stem cells
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(S)-S2.
We show that the rtTA2(S)-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(S)-S2 and neomycin phosphotransferase, we found that embryonic stem cells expressing rtTA2(S)-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.