| Allele Name | transgene insertion R, Jeff W Lichtman |
| Allele Type | Transgenic (Reporter) |
| Allele Synonym(s) | Tg(Thy1-hrGFPII,-mYFP,-tdimer2(12),-mCerulean)R21Lich; Thy1-Brainbow-2.1 line R |
| Gene Symbol and Name | Tg(Thy1-Brainbow2.1)RLich, transgene insertion R, Jeff W Lichtman |
| Gene Synonym(s) | |
| Promoter | Thy1, thymus cell antigen 1, theta, mouse, laboratory |
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| Expressed Gene | CFP, Cyan Fluorescent Protein, jellyfish |
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| Expressed Gene | GFP, Green Fluorescent Protein, |
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| Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
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| Expressed Gene | RFP, Red Fluorescent Protein, coral |
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| Site of Expression | When bred to Cre recombinase expressing mice, there is one of three different inversions for each transgene in each cell of the cre expressing tissue(s). In addition, two excision events may reduce the construct to one of two single invertible DNA segments which can continue to invert as long as cre is present. These different recombination possibilities can be used to express four genes (hrGFPII, mYFP, tdimer2(12) (RFP), or mCerulean (CFP)). Integration of multiple tandem transgene copies allows for labeling of individual neurons and glia in peripheral and central neurons with nuclear localized, membrane-targeted, or cytoplasmic fluorescent proteins in cre recombined cells. |
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| Strain of Origin | C57BL/6J x CBA |
| Chromosome | UN |
| Molecular Note | 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. |
| Mutations Made By | Jeff Lichtman and Joshua Sanes, Harvard University |
