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B6.Cg-Igs7tm94.1(tetO-GCaMP6s)Hze/J
Cat. No.: CEMM-07250526
Common Name: Ai94(TITL-GCaMP6s)-D; Ai94D
Ai94(TITL-GCaMP6s)-D (also called Ai94D) mice have a Cre/Tet-dependent, fluorescent calcium indicator GCaMP6s inserted into 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 GCaMP6 slow variant calcium indicator (GCaMP6s; an ultrasensitive detector of single neuronal action potentials with slower decay and response kinetics). Following subsequent calcium binding (such as neuronal activation), increased EGFP fluorescence is observed.
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| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Igs7tm94.1(tetO-GCaMP6s)Hze
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| Allele Name |
targeted mutation 94.1
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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 |
GCaMP, Genetically encoded calcium indicator
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| MGI Accession ID | show more close |
| Site of Expression |
Expression of fast variant calcium indicator (GCaMP6s) 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 replacement vector was designed with (from 5' to 3') an FRT3 site, two copies of chicken beta-globin HS4 insulator element (to reduce reporter gene expression in absence of transactivators), a modified Tet response element (TRE or tetO), a floxed-STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), the GCaMP6 slow variant calcium indicator coding sequence, 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 GCaMP6 slow variant calcium indicator (GCaMP6s) is an ultrasensitive detector of single neuronal action potentials with slow response kinetics and is an improved version of GCaMP5G. Embryonic stem cells previously targeted with FRT3::AttB::PGK-neoR-polyA::FRT5::splice acceptor::3'hygro cassette::SV40 polyA:AttP in TIGRE, were re-transfected with the pTRE-LSL-GCaMP6s replacement vector and Flp recombinase vector for recombinase-mediated cassette exchange (RCME).Chimeric mice were bred with PhiC31 Gt(ROSA)26Sortm3(phiC31*)Sor mice to remove the AttB/AttP-flanked PGK-hygromycin-SV40polyA cassette and replace it with the recombined AttB/AttP site (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. In 2016, our repository colony is viable and fertile as homozygotes - and we are mainlining our live colony by breeding homozygous mice together.
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For Research Use Only.
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