Tg(Thy1-Brainbow2.1)RLich

The Thy1-Brainbow 2.1 transgene was designed with the mouse Thy1 regulatory elements surrounding four fluorescent proteins (green (GFP), enhanced yellow (EYFP), red (RFP), and cyan (CFP)) sequences uniquely positioned in a tandem fashion and delimited by LoxP sites in opposite orientation. Specifically, this Brainbow 2.1 coding region is composed of two adjacent head to tail tandem dimers, the first contains a LoxP site, humanized Renilla GFP (hrGFPII (Stratagene); with nuclear localization signal), and polyA sequence in forward orientation, and a LoxP site, monomeric EYFP (mYFPA206K), and polyA sequence in reverse orientation. The second head to tail dimer contains a LoxP site, tdimer2(12) RFP, and polyA sequence in forward orientation, and a LoxP site, mCerulean CFP (with membrane tethering palmitoylation sequence), and polyA sequence in reverse orientation. A single FRT site is located at the 3' end of the coding region. The hrGFPII variant of GFP (from Stratagene vector phrGFPII-C) has amino acid substitutions designed to improve spectral properties and performance in mammalian systems. The monomeric EYFP (mYFPA206K) has an amino acid substitution replacing a hydrophobic region with a positively charged residue designed to prevent dimerization. The tdimer2(12) RFP is a non-oligomerizing DsRed variant with a 12 residue linker fusing two copies of the protein (tandem dimer). The monomeric Cerulean (mCerulean) is a variant of ECFP (ECFPS72A/Y145A/H148D/A206K) with amino acid substitutions designed to improve spectral properties and prevent dimerization. (J:125961)