These floxed mutant mice possess loxP sites flanking exon 2 of the Ptger4 gene. This strain may be useful for generating conditional mutations in studies of the role of prostaglandin E2 (PGE2) and function of the PGE2 EP4 receptor.
Richard M. Breyer, Vanderbilt University
Dr. Matthew Breyer, Eli Lilly and Company
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Ptger4 | prostaglandin E receptor 4 (subtype EP4) |
Prostaglandin E2 (PGE2), a fatty acid derivative and a primary target of NSAIDs, is important in diverse physiological processes, including but not limited to: contraction and relaxation of smooth muscle, vasodilation, vasoconstriction, blood pressure regulation, immune response and inflammation regulation and bone formation and healing. There are four receptors for PGE2. The Ptger4 gene encodes a G-protein coupled PGE2 receptor (EP4), which is essential for embryonic development and neonatal survival in mice.
These mice possess loxP sites on either side of exon 2 of the targeted gene. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues. Removal of the floxed sequence creates a null allele. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
When bred to a strain with Cre recombinase expression in the myeloid lineage, this mutant mouse strain may be useful in studies of innate immunity and inflammation in the CNS.
A targeting vector designed by Dr. Matthew Breyer (Vanderbilt University) containing a loxP site flanked PGK-Neo selection cassette was utilized in the construction of this mutant. This selection cassette was inserted downstream of exon 2 of the targeted gene, and another loxP site was inserted upstream of exon 2. This construct was electroporated into 129S6/SvEvTac derived TL-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts.
The resulting male EP4lox+neo chimeric animals were bred to C57BL/6 females. Heterozygous EP4lox+neo mice were intercrossed to generate homozygotes. Mice homozygous for the EP4lox+neo allele die perinatally due to patent ductus arteriosus. Heterozygous EP4lox+neo male mice were then bred to B6D2 F1 females to produce fertilized oocytes, which were subsequently transfected with a Cre recombinase plasmid. The resulting single cell embryos were transferred to pseudopregnant females. The male offspring that was mosaic for all 3 alleles was bred to C57BL/6 females to produce heterozygous EP4flox mice. Heterozygous EP4flox were intercrossed to obtain homozygotes. The donating investigator reported that mice were then backcrossed to C57BL/6J for 10 generations (see SNP note below).
During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 27 markers throughout the genome was segregating with 129 or DBA on Chromosome 15. Also, 4 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
Allele Name | targeted mutation 1.1, Matthew D Breyer |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | EP4flox |
Gene Symbol and Name | Ptger4, prostaglandin E receptor 4 (subtype EP4) |
Gene Synonym(s) | |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 15 |
Molecular Note | The neo cassette was removed via partial cre recombination leaving loxP sites surrounding exon 2, which encodes slightly more than five of the seven transmembrane-spanning domains. Recombination in mutant mice was confirmed by Southern blot and PCR. |
When maintaining a live colony, these mice can be bred as homozygotes.
When using the EP4flox mouse strain in a publication, please cite the originating article(s) and include JAX stock #028102 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Ptger4<tm1.1Matb> |
Frozen Mouse Embryo | B6.129S6(D2)-Ptger4<tm1.1Matb>/BreyJ Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.129S6(D2)-Ptger4<tm1.1Matb>/BreyJ Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.129S6(D2)-Ptger4<tm1.1Matb>/BreyJ Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6.129S6(D2)-Ptger4<tm1.1Matb>/BreyJ Frozen Embryo | $3373.50 |
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