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B6.Cg-Igs7tm210(tetO-GCaMP7f, CAG-tTA2)Tasic/J
Cat. No.: CEMM-07251106
Common Name: TITL-GC7f-ICF-IRES-tTA2
Ai210 (also called Ai210 (TITL-GC7f-ICF-IRES-tTA2)) is a dual-recombinase responsive (cre/flp-dependent), Tet-controllable, fluorescent calcium-indicator tool allele created by targeted insertion at the Igs7 locus (TIGRE). Exposure to both Cre and FLP recombinase results in expression of tTA2 and jGCaMP7f (an ultrasensitive EGFP fluorescent calcium detector of neuronal activity with faster decay and response kinetics) in those cells where cre and FLP expression overlaps/intersects. The jGCaMP7f expression may be expected to be diminished/eliminated by doxycycline. jGCaMP7f has high sensitivity and affinity, improved detection of individual spikes and may allow tracking of larger populations of neurons using two-photon imaging.
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| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Igs7tm210(tetO-GCaMP7f,CAG-tTA2)Tasic
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| Allele Name |
targeted mutation 210
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| Allele Attributes |
Conditional ready (e.g. floxed); Reporter; Inducible; Transactivator; Epitope tag
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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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| MGI Accession ID | show more close |
| Strain of Origin |
(129S6/SvEvTac x C57BL/6NCrl)F1
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| Molecular Note |
The targeting vector contains (from 5' to 3') a Bxb1 attL site, a partial green fluorescent protein sequence (non-functional 500bp fragment to inCrease the space between the Bxb1 attL site and the insulator sequence to improve stability of the targeting vector), two copies of chicken beta-globin HS4 insulator element (to reduce reporter gene expression in absence of transactivators), a Tet response element/promoter (tetO), a loxP-flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-PGKpA), a GCaMP7 sensitive and fast variant calcium indicator sequence (jGCaMP7f; details below) followed by a 6xHis tag, T7 tag and XPress tag sequence, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA, two copies of chicken beta-globin HS4 insulator element, a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG; from pCAGGS), a FRT flanked STOP cassette (stop codons in all three reading frames linked to synthetic pA-hGHpA-TKpA), an internal ribosome entry site (IRES) sequence, a synthetic modified tetracycline-regulated transactivator gene (tTA2(S)), a WPRE, a BGH polyA, a PhiC31 attB site, a PGK-5'hygro cassette, an mRNA splice donor, a Bxb1 attR site, SA, 3'hygro, SV40pA, and PhiC31 attP. The allele carries a chicken smooth muscle M13 fragment of myosin light chain kinase (also called calmodulin-binding peptide [CBP]), a circularly permutated EGFP (cpEGFP; aa 149-238 followed by aa 1-144 [with mutations to inCrease dynamic range/baseline fluorescence, as well as inCreased sensitivity/slower kinetics]), and a rat calmodulin DNA fragment (CaM; aa 2-148 [with mutations to inCrease the fluorescence change for small calcium transients]). Amino acid substitutions in the cpEGFP-to-CaM linker improve sensor function. In addition, this allele has the H78K mutation in GFP and the A317L, R381T, T383S, and G392R mutations in CaM.
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| HUSBANDRY | |
|---|---|
| Suggested Controls |
C57BL/6J
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| Breeding Considerations |
Heterozygous mice are viable and fertile with no reported gross physical or behavioral abnormalities. attempts have not been made to make this strain homozygous. When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice.
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| Breeding Strategy |
Heterozygote x Heterozygote
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For Research Use Only.
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