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B6.Cg-Igs7tm92.1(tetO-ECFP*/Venus*)Hze/J
Cat. No.: CEMM-07250594
Common Name: Ai92(TITL-YCX2.60)-D; Ai92D
Ai92(TITL-YCX2.60)-D (also called Ai92D) mice are a Cre/Tet-dependent, fluorescent calcium indicator line, created by targeted insertion of floxed-STOP-YCX2.60 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 cassettes by Cre recombinase, they may be used to generate Tet-Off/Tet-On mutant animals with conditional (inducible/reversible) expression of a FRET-based Ca2+ sensor with expanded dynamic range (YCX2.60). Following subsequent calcium binding (such as neuronal activation), robust yellow fluorescence and diminished cyan 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 |
Igs7tm92.1(tetO-ECFP*/Venus*)Hze
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| Allele Name |
targeted mutation 92.1
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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 |
CFP, Cyan Fluorescent Protein, jellyfish; YFP, Yellow Fluorescent Protein, jellyfish
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| MGI Accession ID | show more close |
| Site of Expression |
Expression of yellow fluorescent protein and cyan fluorescent protein 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 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 YCX2.60 coding sequence [an enhanced cyan fluorescent protein with C-terminal 11 amino acids deleted (ECFPDeltaC11 with R27K and K213N mutations, a rat calmodulin DNA fragment (CaM; aa 5-148 with N60D and D78Y mutations [to inCrease the fluorescence change for small calcium transients] and E127V mutation), a GG linker, a chicken smooth muscle M13 fragment of myosin light chain kinase (with I-to-S mutation in the 20th aa of the sequence), and a circularly permutated Venus enhanced yellow fluorescent protein (cp173 Venus; aa 174-239, a 15 bp linker and aa 1-173)], a WPRE sequence, 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 fluorescence resonance energy transfer (FRET)-based calcium sensor Yellow Cameleon variant X 2.60 (YCX2.60) is an improved version of YC2.60 with expanded dynamic range and 2-3-fold greater dissociation constant. The fusion gene is an enhanced cyan fluorescent protein with C-terminal 11 amino acids deleted, a rat calmodulin DNA fragment (CaM; aa 5-148 with N60D and D78Y mutations [to inCrease the fluorescence change for small calcium transients] and E127V mutation), a GG linker, a chicken smooth muscle M13 fragment of myosin light chain kinase (with I-to-S mutation in the 20th aa of the sequence), and a circularly permutated Venus enhanced yellow fluorescent protein (cp173 Venus; aa 174-239, a 15 bp linker and aa 1-173). 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 | |
|---|---|
| Suggested Controls |
C57BL/6J Wild-type from the colony
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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. The donating investigator has not attempted to generate homozygous mice to date.
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For Research Use Only.
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