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B6;129S-Igs7tm85.1(tetO-gltI/GFP*)Hze/J
Cat. No.: CEMM-07250593
Common Name: Ai85(TITL-iGluSnFr)-D
Ai85(TITL-iGluSnFr)-D (also called Ai85D) mice are a Cre/Tet-dependent, fluorescent glutamate indicator line, created by targeted insertion of floxed-STOP-iGluSnFr at the Igs7 locus (TIGRE; an intergenic region on mouse chromosome 9 that allows reporter expression to be tightly regulated). After removal of the floxed-STOP cassette by Cre recombinase, they may be used to generate Tet-Off/Tet-On mutant animals with conditional (inducible/reversible) expression of iGluSnFr, an intensity-based glutamate-sensing fluorescent reporter that is fast and sensitive. Increased GFP fluorescence is observed following subsequent glutamate binding (such as excitatory synaptic activity).
Inquiry
| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Igs7tm85.1(tetO-gltI/GFP*)Hze
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| Allele Name |
targeted mutation 85.1
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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 |
GFP, Green Fluorescent Protein; myc tag
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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 pTRE-LSL-iGluSnFr replacement vector was designed with (from 5' to 3') an FRT3 site, two copies of chicken beta-globin HS4 insulator element, a modified Tet response element promoter, a floxed-STOP cassette, the iGluSnFr coding sequence (details below), a WPRE sequence (to enhance mRNA stability), a BGH polyA, two copies of chicken beta-globin HS4 insulator element, an AttB site, a PGK-5'hygro cassette, an RNA splice donor and a FRT5 site. The iGluSnFr fusion gene is constructed from the Escherichia coli gltI gene and circularly permutated green fluorescent protein (cpGFP). Specifically, iGluSnFr has a 21 aa IgG K-leader sequence, the gltI aa 1-253, the LILV linker, the cpGFP (aa 148-239, an 18 bp flexible linker and aa 1-147), the L2NP linker, the gltI aa 254-279, an 11 aa c-Myc epitope and a 49 aa PDGR sequence (PDGR aa 513-561). iGluSnFr is an extremely fast and sensitive glutamate indicator with large signal-to-noise ratio, and kinetics appropriate for in vivo imaging. PhiC31-mediated recombination removed the AttB/AttP-flanked PGK-hygromycin-SV40polyA cassette and replaced it with the recombined AttB/AttP site (AttL).
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| HUSBANDRY | |
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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. Alternatively, homozygous mice may be bred together. In 2017, breeding homozygous mice together successfully identified that homozygous Ai85D mice are viable and fertile.
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For Research Use Only.
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