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B6.Cg-Igs7tm94.1(tetO-GCaMP6s)Hze Tg(Camk2a-tTA)1Mmay/J
Cat. No.: CEMM-07250533
Common Name: Ai94(TITL-GCaMP6s)-D; CaMK2a-tTA
Ai94(TITL-GCaMP6s)-D;CaMK2a-tTA (or Ai94D;CaMK2a-tTA) 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), 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 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 in those cells. 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. Founder line 1 has >20 transgene copies.
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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, mice heterozygous or homozygous for the Ai94D targeted mutation (Igs7tm94.1(tetO-GCaMP6s)Hze) and hemizygous for the CaMK2a-tTA transgene (Tg(Camk2a-tTA)1Mmay) may be bred to mice heterozygous or homozygous for Ai94D and wildtype (non-carrier) for CaMK2a-tTA. Ai94D;CaMK2a-tTA mice heterozygous or homozygous for Ai94D and hemizygous for CaMK2a-tTA are viable and fertile with no reported gross physical or behavioral abnormalities. it has not been attempted to make Ai94D;CaMK2a-tTA animals homozygous for the CaMK2a-tTA transgene.Regarding viability and fertility of the single mutant parental mouse lines: For the single mutant parental mouse line for Ai94D, the donating investigator reports that mice homozygous for Ai94D are viable and fertile with no reported gross physical or behavioral abnormalities. Similarly in 2016, Repository is viable and fertile as homozygotes. For the CaMK2a-tTA parental mouse line, it has not been attempted to make the CaMK2a-tTA transgenic mouse line homozygous to date.
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For Research Use Only.
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