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B6.Cg-Igs7tm133.1(tetO-apx1*)Hze/J
Cat. No.: CEMM-07250784
Common Name: Ai133(TITL-ssAPEX2tm)-D (or Ai133D)
Ai133(TITL-ssAPEX2tm)-D (also called Ai133D) mice are a Cre/Tet-dependent, substrate-dependent reporter line - created by targeted insertion of floxed-STOP-APEX2 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 cassette by Cre recombinase, they may be used to generate Tet-Off/Tet-On mutant animals with conditional (inducible/reversible) expression of APEX2 - an engineered peroxidase with improved catalytic efficiency that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics.
Inquiry
| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Igs7tm133.1(tetO-apx1*)Hze
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| Allele Name |
targeted mutation 1.1
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| Allele Attributes |
Conditional ready (e.g. floxed); Inducible; Inserted expressed sequence
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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 |
| Site of Expression |
APEX2 is expressed in cells/tissues where the expression patterns of the individual promoters driving Cre and tTA/rtTA overlap, and expression can be suppressed with tetracycline or its analog doxycycline (dox).
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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, a modified Tet response element promoter, a floxed-STOP cassette, the ssAPEX2tm coding sequence (details below), 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. 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-ssAPEX2tm replacement vector and Flp recombinase vector for recombinase-mediated cassette exchange (RCME). PhiC31-mediated recombination removed the AttB/AttP-flanked sequence (PGK-Neo-polyA::frt5::RNA splice acceptor::3'hygro-polyA) and replaced it with the recombined AttB/AttP site (AttL). APEX2 is a second-generation variant of soybean Glycine hispida ascorbate peroxidase gene (apx1) that harbors the three APEX mutations (K14D, W41F, E112K) optimized for expression in mice, as well as one additional mutation (A134P) for improved catalytic efficiency. The ssAPEX2tm fusion gene has APEX2 fused to a T-cell Cd2 antigen signal sequence (ss) and transmembrane targeting sequence (tm). There is a FLAG-tag domain (DYKDDDDK) between the signal sequence and the APEX2 sequence.
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| HUSBANDRY | |
|---|---|
| Suggested Controls |
C57BL/6J
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| Breeding Considerations |
The donating investigator reports that heterozygous and homozygous mice are viable and fertile with no reported gross physical or behavioral abnormalities. When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice. Alternatively, homozygous mice may be bred together.
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| Breeding Strategy |
Homozygote x Homozygote
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For Research Use Only.
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