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B6.Cg-Igs7tm90.1(tetO-COP4*/EGFP)Hze/J
Cat. No.: CEMM-07250523
Common Name: Ai90(TITL-Chronos)-D; Ai90D
Ai90(TITL-Chronos)-D (also called Ai90D) mice are a Cre/Tet-dependent, optogenetic line, created by targeted insertion at the Igs7 locus (TIGRE; an intergenic region on mouse chromosome 9 that allows reporter expression to be tightly regulated). Following Cre-mediated removal of the STOP cassette, they may be used to generate Tet-Off/Tet-On mutant animals with conditional (inducible/reversible) expression of the channelrhodopsin/EGFP fusion protein (Chronos/EGFP). Subsequent illumination of Chronos-expressing (EGFP fluorescent) cells with blue-to-green light leads to reversible photostimulation of action potential firing/neural activity in these cells.
Inquiry
| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Igs7tm90.1(tetO-COP4*/EGFP)Hze
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| Allele Name |
targeted mutation 90.1
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| Allele Attributes |
Conditional ready (e.g. floxed); Reporter; Inducible
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| Gene Symbol |
Igs7
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| Gene Name |
intergenic site 7
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| Chromosome |
9
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| Expressed Genes |
COP4, Channelrhodopsin, Chlamydomonas; GFP, Green Fluorescent Protein
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| MGI Accession ID | show more close |
| Site of Expression |
Expression of GFP is under the control of tetO in cells/tissues where promoters driving Cre recombinase are expressed.
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| Strain of Origin |
(129S6/SvEvTac x C57BL/6NCrl)F1
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| Molecular Note |
The targeting vector was designed with (from 5' to 3') an FRT3 site, two tandem copies of the chicken beta-globin HS4 insulator element (to reduce reporter gene expression in absence of transactivator protein), a modified Tet response element (TRE or tetO), a loxP-flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), the COP4*/EGFP coding sequence, a WPRE sequence (to enhance mRNA transcript stability), a bovine growth hormone polyA signal, two tandem copies of the chicken beta-globin HS4 insulator element, an AttB site, a PGK-5' hygro cassette, an RNA splice donor and an FRT5 site.The COP4*/EGFP fusion protein is composed of a mammalian codon-optimized Stigeoclonium helveticum-derived channelrhodopsin ShChR (Chronos) fused in-frame via a 18 nucleotide linker to the N-terminus of enhanced green fluorescent protein (EGFP). Chronos is a blue-to-green light-driven (~470-530 nm) cation channel that depolarizes the cell and causes action potentials. Chronos has high light sensitivity and faster kinetics than previous channelrhodopsins. Embryonic stem cells, previously targeted with FRT3-AttB-PGK-neoR-polyA::FRT5::splice acceptor::3' hygro cassette::SV40 polyA: AttP in the same locus, were re-transfected with the targeting vector and Flp recombinase vector. Mutant mice were bred with Gt(ROSA)26Sortm3(phiC31*)Sor mice to remove to remove the AttB/AttP-flanked PGK-hygro-SV40pA cassette and replace it with the recombined AttB/AttP site (AttL). The Ai90D allele is therefore TIGRE-frt3-Ins-TRE-LSL-Chronos/EGFP-WPRE-bGHpA-Ins-AttL.
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| HUSBANDRY | |
|---|---|
| Suggested Controls |
C57BL/6J
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| Breeding Considerations |
When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or to C57BL/6J inbred mice. The donating investigator has not attempted to generate homozygous mice to date.
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For Research Use Only.
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