Tet-On Binary Systems for Tissue-Specific and Inducible Transgene Expression

Abstract

Tissue-specific and inducible control of transgene expression is a cornerstone of modern studies in developmental biology. Even though such control of transgene expression has been accomplished in Xenopus, no general or widely available set of transgenic lines have been produced akin to those found in mouse and zebrafish. Here, I describe the design and characterization of transgenic lines in Xenopus constituting the Tet-On binary transgene expression system comprising two components: (1) rtTA transgenic lines, i.e., lines harboring the doxycycline- (Dox-) dependent transgenic transcription factor rtTA under control of a tissue-specific promoter and (2) transgenic promoter (TRE) transgenic lines, i.e., lines harboring a gene of interest (hereafter called the transgene) under control of a promoter (TRE). In double transgenic animals, i.e., embryos or tadpoles harboring both the rtTA and TRE components, transgene expression remains off the absence of Dox. Addition of Dox to the rearing water causes a conformational change in rtTA allowing it to bind the TRE promoter and induce transgene expression. Tissue specificity of transgene expression is determined by the promoter regulating rtTA expression, and inducibility is determined by the addition of Dox to the rearing water. Deposition of rtTA and TRE transgenic lines enabling tissue-specific inducible control of transgene expression into the Xenopus stock center will provide a powerful and flexible resource for studies in developmental biology.