Gt(ROSA)26Sor^{tm2.1(CAG-rtTS)Gev}

Overlap extension PCR was used to clone the tetracycline responsive domain from rtTA*M2 (Clontech, pTet-on Advanced) to the KRAB-AB silencing domain of the Kid-1 protein (SDKid-1) (Clontech, pTet-tTS), creating the chimeric rtTS protein (tetracycline-controlled trans-silencer). The SA70b pROSA-CAGGS-attP50/B53-hygro-NLSLacZ vector was obtained from ARTEMIS Pharmaceuticals. A fragment containing the CAGGS promoter and an intron was removed from SA70b and cloned into a modified pROSA26 plasmid48, creating the pROSA26CAG-PAS vector. The rtTS cDNA above was cloned into pBigT, and then into pROSA26CAG-PAS to create the final Rosa26 CAG-LSL-rtTS targeting vector. Cre-mediated recombination removed the lox-stop-lox cassette. (J:255811)