In this strain the entire coding region of the Cldn6 gene is replaced with a CreERT2 fusion protein, an internal ribosome entry site (IRES) and a histone H2B-Venus fluorescent protein. These mice may be useful for visualizing endoderm specification, proliferation, and patterning in the developing embryo.
Douglas A Melton, Harvard University
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing, Inducible) | Cldn6 | claudin 6 |
In this strain, the Cldn6CIHV allele replaces the entire coding region of the claudin 6 (Cldn6) locus with a CreERT2 fusion protein, an internal ribosome entry site (IRES), and a histone H2B-Venus fluorescent protein. This abolishes gene expression. Homozygotes are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. CLDN6 is a structural protein involved in tight junction formation, with a functional role in the epidermal permeability barrier. In this strain endogenous Cldn6 promoter/enhancer regions drive Cre-ERT2 expression and Venus immunofluorescence in embryonic endoderm during organogenesis. Cre-ERT2 fusion gene activity is inducible and only observed following tamoxifen administration. When Cldn6CIHV mice are bred with mice containing loxP-flanked sequence, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Cldn6-expressing cells of the offspring. These mice may be useful for visualizing endoderm specification, proliferation, and patterning in the developing embryo.
The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, Cre-ERT2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered.
A targeting vector was designed to replace the entire coding sequence of the claudin 6 (Cldn6) gene with, from 5' to 3', a CreERT2 fusion protein, an internal ribosome entry site (IRES), a histone H2B-Venus fluorescent protein, an SV40 polyadenylation signal, and a self-excising ACN neomycin (neo) selection cassette. This construct was electroporated into 129X1/SvJ-derived AV3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6J females. These Cldn6CIHV mutant mice were maintained on this mixed B6;129X1 genetic background. Upon arrival at The Jackson Laboratory, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
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Site of Expression | Cre-ERT2 expression and Venus immunofluorescence is seen in embryonic endoderm during organogenesis. |
Allele Name | targeted mutation 1, Douglas A Melton |
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Allele Type | Targeted (Recombinase-expressing, Inducible) |
Allele Synonym(s) | Cldn6CIHV; Cldn6tm1(cre/ESR1)Dam |
Gene Symbol and Name | Cldn6, claudin 6 |
Gene Synonym(s) | |
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | Cre-ERT2 expression and Venus immunofluorescence is seen in embryonic endoderm during organogenesis. |
Strain of Origin | 129X1/SvJ |
Chromosome | 17 |
Molecular Note | The entire reading frame contained within exon 3 was replaced with the CIHV cassette that contains cre-ER |
Mutations Made By | Douglas Melton, Harvard University |
When maintaining a live colony, homozygous mice may be bred together.
When using the B6;129X1-Cldn6tm1(cre/ERT2)Dam/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #014638 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Cldn6<tm1(cre/ERT2)Dam> |
Frozen Mouse Embryo | B6;129X1-Cldn6<tm1(cre/ERT2)Dam>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129X1-Cldn6<tm1(cre/ERT2)Dam>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129X1-Cldn6<tm1(cre/ERT2)Dam>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129X1-Cldn6<tm1(cre/ERT2)Dam>/J Frozen Embryo | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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