Gt(ROSA)26Sor^{tm1.1(CAG-Pycard/mCitrine*,-CD2*)Dtg}

The targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, a loxP-flanked STOP cassette, the mASC-citrine-HA fusion protein (described below), an FRT site, an iIRES, a portion from the human CD2 antigen locus encoding a truncated signaling-deficient protein (hCD2*), a second FRT site and a BGH polyA signal. The targeting vector was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The mASC-citrine-HA fusion protein cDNA sequence encodes the mouse PYD and CARD domain containing coding region fused to a Citrine variant of yellow fluorescent protein, all followed by a hemagglutinin tag sequence. The Citrine used here is a yellow fluorescent protein variant with the K69A and A206K mutations (for improved brightness and monomeric stability) - encoding a ~5.7 pKa monomer with ~488nm excitation and ~529nm emmission spectra. Cre-mediated recombination removed the upstream loxP-flanked STOP cassette. (J:238511)