Sema3dGFPcre knock-in/knock-out mice express a GFPcre fusion protein under the direction of the Sema3d promoter. They may be useful for cellular fate mapping during pulmonary vein formation.
Jonathan A Epstein, University of Pennsylvania
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing) | Sema3d | sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3D |
Sema3dGFPcre mice express a GFPcre fusion protein from the endogenous sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3D (Sema3d) promoter. The presence of GFPcre abolishes expression of Sema3d. SEMA3D is a secreted guidance molecule crucial for the normal patterning of pulmonary veins. Sema3d cellular derivatives populate the endothelial lining of the sinus venosus (SV) by E10.5. Expression is restricted, in the heart, to only proepicardial cells and migrating epicardial cells prior to E11.5, with strong epicardial expression persisting beyond E14.5. Homozygous males are mostly viable and fertile, while homozygous females do not breed. Heterozygous mice are viable and fertile. Sema3deGFPcre adult mice show abnormalities of pulmonary venous patterning, including massively enlarged right atria and ventricles.
When these mice are bred with mice containing loxP-flanked sequence, Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing cells of the offspring.
A targeting construct was designed to replace the initiation codon in exon 2 of the sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3D (Sema3d) gene with an enhanced green fluorescent protein fused to a Cre Recombinase sequence (GFPcre), followed by a frt-flanked neomycin resistance (neo) cassette. The construct also replaced the remaining downstream sequence of exon 2. This construct was electroporated into 129S6/SvEvTac-derived TL1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric animals were crossed to Tg(ACTFLPe)9205Dym mice on a mixed background to remove the neo selection cassette. These Sema3deGFPcre mice were bred together to remove the Flp-expressing transgene and were subsequently bred to C57BL/6 mice for at least 1 generation. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.
Expressed Gene | GFP/cre, Green Fluorescent Protein/Cre recombinase fusion protein, |
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Site of Expression | Expression is restricted in the heart to proepicardial cells and migrating epicardial cells. |
Allele Name | targeted mutation 2.1, Jonthan Epstein |
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Allele Type | Targeted (Recombinase-expressing) |
Allele Synonym(s) | Sema3DGFPcre; Sema3dtm1.1(GFP/cre)Cjt |
Gene Symbol and Name | Sema3d, sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3D |
Gene Synonym(s) | |
Expressed Gene | GFP/cre, Green Fluorescent Protein/Cre recombinase fusion protein, |
Site of Expression | Expression is restricted in the heart to proepicardial cells and migrating epicardial cells. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 5 |
Molecular Note | An eGFP/cre fusion gene and an Frt-flanked neo cassette was inserted into the Sema3d locus at the ATG codon, replacing the remaining exon 2 sequence by homologous recombination in TL-1 ES cells. The neo was deleted by crossing germline mutants to Flp-expressing mice. |
When maintaining a live colony heterozygous females may be bred to homozygous males. The donating investigator reports that homozygous males are mostly viable and fertile, while homozygous females do not breed.
When using the Sema3DGFPcre mouse strain in a publication, please cite the originating article(s) and include JAX stock #029270 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Sema3d<tm2.1(GFP/cre)Joe> |
Frozen Mouse Embryo | STOCK Sema3d<tm2.1(GFP/cre)Joe>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | STOCK Sema3d<tm2.1(GFP/cre)Joe>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | STOCK Sema3d<tm2.1(GFP/cre)Joe>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | STOCK Sema3d<tm2.1(GFP/cre)Joe>/J Frozen Embryo | $3373.50 |
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