HCN4CreERT2 mice express CreERT2 in the first heart field of differentiating myocytes in the developing cardiac crescent.
Sylvia M Evans, University of California, San Diego
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing, Inducible) | Hcn4 | hyperpolarization-activated, cyclic nucleotide-gated K+ 4 |
Heterozygous mice are viable and fertile, homozygotes are embryonic lethal. The HCN4CreERT2 allele expresses CreERT2 from the hyperpolarization-activated, cyclic nucleotide-gated K+ 4 (HCN4) promoter/enhancer elements. HCN4 is expressed in the first heart field of differentiating myocytes in the developing cardiac crescent. Cre-ERT2 fusion gene activity is observed in the same cells only following tamoxifen administration. When these mice are bred with mice containing loxP-flanked sequence, Cre-mediated recombination will result in deletion of the floxed sequences in the cardiac conduction system (CCS) of the offspring.
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 insert a CreERT2 coding sequence and frt-flanked neomycin resistance selection cassette immediately before endogenous start codon of hyperpolarization-activated, cyclic nucleotide-gated K+ 4 (HCN4) gene. This construct was electroporated into embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred. Mice were bred to Flp-transgenic mice on a Black Swiss background to remove the neo cassette, and progeny were crossed to remove the Flp transgene. These mice were maintained on a Black Swiss outbred 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 | Mice carrying this allele express CreERT2 in the first heart field of differentiating myocytes in the developing cardiac crescent. |
Allele Name | targeted mutation 2.1, Sylvia Evans |
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Allele Type | Targeted (Recombinase-expressing, Inducible) |
Allele Synonym(s) | HCN4CreERT2 |
Gene Symbol and Name | Hcn4, hyperpolarization-activated, cyclic nucleotide-gated K+ 4 |
Gene Synonym(s) | |
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | Mice carrying this allele express CreERT2 in the first heart field of differentiating myocytes in the developing cardiac crescent. |
Strain of Origin | Not Specified |
Chromosome | 9 |
Molecular Note | A targeting vector was designed to insert a CreERT2 coding sequence and frt-flanked neomycin resistance selection cassette immediately before endogenous start codon. The neo cassette was removed by FLP mediated recombination. |
When maintaining a live colony, heterozygous mice may be bred to wildtype (noncarrier) mice from the colony. Homozygotes are embryonic lethal.
When using the HCN4CreERT2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #024283 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Hcn4<tm2.1(cre/ERT2)Sev> |
Frozen Mouse Embryo | STOCK Hcn4<tm2.1(cre/ERT2)Sev>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | STOCK Hcn4<tm2.1(cre/ERT2)Sev>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | STOCK Hcn4<tm2.1(cre/ERT2)Sev>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | STOCK Hcn4<tm2.1(cre/ERT2)Sev>/J Frozen Embryo | $3373.50 |
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