New doxycycline-inducible transgenic lines in Xenopus.

We have characterized two new transgenic Xenopus lines enabling transgene expression using the Tet-On inducible system. An inducer line expresses the doxycycline- (Dox-) activated transcription factor rtTA under control of the ubiquitous promoter CMV. A responder line enables Dox-inducible expression of a dominant positive thyroid hormone receptor via a tetracycline responsive transgenic promoter (TRE). Dox-induced expression of transgenic GFP mRNA was detectable after 3 hr and increased up to 10- to 50-fold by 2 days depending on dose of Dox. Induced GFP mRNA expression returned to uninduced levels within 3 days upon Dox removal. Treatment of rtTA inducer and TRE responder double transgenic animals with Dox caused acceleration of metamorphic changes in thyroid hormone-response gene expression and morphology. These transgenic lines will be made available through the new Xenopus Stock Center and will serve as valuable tools for genetic analysis of development and metamorphosis.