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STOCK Igs7tm93.1(tetO-GCaMP6f)Hze Tg(Camk2a-tTA)1Mmay/J
Cat. No.: CEMM-07250530
Common Name: Ai93(TITL-GCaMP6f)-D; CaMK2a-tTA
Ai93(TITL-GCaMP6f)-D;CaMK2a-tTA (or Ai93D;CaMK2a-tTA) mice have a Cre/Tet-dependent, fluorescent calcium indicator GCaMP6f inserted into the Igs7 locus (TIGRE; an intergenic region on mouse chromosome 9 that allows reporter expression to be tightly regulated), as well as a transgene directing tetracycline-controlled transactivator protein (tTA) expression in forebrain neurons. After removal of the floxed-STOP cassette by Cre recombinase, resulting mice have doxycycline-inducible/reversible expression of GCaMP6 fast variant calcium indicator (GCaMP6f; a detector of single neuronal action potentials with fast response kinetics) in forebrain neurons. Following subsequent calcium binding (such as neuronal activation), bright EGFP fluorescence is observed. As described for, the CaMK2a-tTA transgene integrated into chromosome 12 causing a 508 Kb deletion that spans the 3' half of Vipr2, the entire Wdr60, Esyt2, D430020J02Rik and Ncapg2 loci and the first two exons of Ptprn2. Homozygous mice will therefore have a functional knock-out of the deleted loci, and altered or null expression of Vipr2 and Ptprn2.
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| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Igs7tm93.1(tetO-GCaMP6f)Hze
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| Allele Name |
targeted mutation 93
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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 (GCaMP6f) 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 GCaMP6 fast variant calcium indicator (GCaMP6f) coding sequence, a WPRE sequence (to enhance the 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 siteThe GCaMP6 fast variant calcium indicator (GCaMP6f) is a detector of single neuronal action potentials with fast response kinetics, and is an improved version of GCaMP5G. GCaMP6f has a chicken smooth muscle M13 fragment of myosin light chain kinase, a circularly permutated EGFP, and a rat calmodulin DNA fragment mutation for accelerated kinetics. Embryonic stem cells, previously targeted with FRT5-AttB-Hygro2 cassette-AttP-SV40pA-splice acceptor-PGKpA-FRT3 in the same locus, were re-transfected with the targeting vector and Flp recombinase vector. Genotyping/sequencing revealed that Ai93 had two tandem copies of the pTRE-LSL-GCaMP6f replacement vector inserted during RCME. Chimeric mice were bred with PhiC31 Gt(ROSA)26Sortm3(phiC31*)Sor mice to remove the AttB/AttP-flanked region. The Ai93D allele is therefore TIGRE-frt3-Ins-TRE-LSL-GCaMP6f-WPRE-bGHpA-Ins-AttB-PGK-5'hygro-SD-frt5-frt3-Ins-TRE-LSL-GCaMP6f-WPRE-bGHpA-Ins-AttL.
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| HUSBANDRY | |
|---|---|
| Suggested Controls |
B6129SF2/J
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| Breeding Considerations |
When maintaining a live colony, mice heterozygous for the Ai93D targeted mutation (Igs7tm93.1(tetO-GCaMP6f)Hze) and hemizygous for the CaMK2a-tTA transgene may be bred together. Alternatively, mice heterozygous or homozygous for Ai93D and wildtype (non-carrier) for CaMK2a-tTA may be bred with mice wildtype for Ai93D and hemizygous or homozygous for CaMK2a-tTA. The phenotype of mice homozygous for both Ai93D and CaMK2a-tTA has not been characterized to date.
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| Breeding Strategy |
See "Breeding Considerations"
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For Research Use Only.
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