These Prg4GFPCreERt2 mice express the inducible GFP/cre/ERT2 fusion gene in a pattern that mimics the expression of the endogenous Prg4 gene: in the chondrocytes of articular cartilage but not in the growth plate chondrocytes.
Matthew Warman, Boston Children's Hospital, Harvard Medical School
Andrew B. Lassar, Harvard Medical School
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Recombinase-expressing, Inducible) | Prg4 | proteoglycan 4 (megakaryocyte stimulating factor, articular superficial zone protein) |
The Prg4, proteoglycan 4 (megakaryocyte stimulating factor, articular superficial zone protein), gene encodes the lubricin protein, a large proteoglycan that acts as a lubricant at the surface of cartilage of articulating joints. Prg4 also encodes for the superficial zone protein, megakaryocyte stimulating factor, and hemangiopoietin. These mice carry a targeted mutation (knock in) allele in which a GFP/cre/ERT2 fusion gene has been inserted into exon 2 of the Prg4 gene. Mice that are homozygous for the targeted mutation are viable and fertile, but exhibit a loss of the superficial-most layer of articular chondrocytes and displayed synoviocyte hyperplasia in joints. Expression of the GFP/cre/ERT2 fusion gene mimics expression of the endogenous Prg4 gene.
When these Prg4GFPCreERt2 mice are bred with mice containing a loxP flanked sequence of interest, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the flanked sequences in Prg4 expressing cells.
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 designed in the laboratory of Dr. Andrew B. Lassar (Harvard Medical School) containing a GFPCreERt2, Green Fluorescent Protein, Cre recombinase and estrogen receptor 1 (human) fusion gene and an FRT flanked PGK-NEO selection cassette, was inserted into the translation initiation codon of exon 2 of the Prg4, proteoglycan 4 (megakaryocyte stimulating factor, articular superficial zone protein). The construct was electroporated into unspecified 129/SV derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were crossed to FLP recombinase expressing mice (ACT-FLPe) mice (on an unknown genetic background) to remove the FRT flanked PGK-NEO selection cassette. The mice were crossed to mice carrying the Gt(ROSA)26Sortm1Sor allele (Stock No. 003474). Further breeding of the double mutant male mice to C57BL/6 female mice was completed to remove the Gt(ROSA)26Sortm1Sor allele.
Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | GFP/cre/ERT2 fusion protein is expressed in the top layer cells of the articular cartilage (in younger mice) or in the top and deeper layers of articular cartilage (in older mice). |
Allele Name | targeted mutation 1, Andrew Lassar |
---|---|
Allele Type | Targeted (Recombinase-expressing, Inducible) |
Allele Synonym(s) | Prg4GFPCreERt2 |
Gene Symbol and Name | Prg4, proteoglycan 4 (megakaryocyte stimulating factor, articular superficial zone protein) |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | GFP/cre/ERT2 fusion protein is expressed in the top layer cells of the articular cartilage (in younger mice) or in the top and deeper layers of articular cartilage (in older mice). |
Strain of Origin | 129 |
Chromosome | 1 |
General Note | ES cells = unspecified 129/SV |
Molecular Note | A targeting vector containing a GFP/cre/ERT2 (green fluorescent protein, cre recombinase and estrogen receptor 1 tamoxifen binding domain fusion gene and an FRT flanked PGK-NEO selection cassette), was inserted into the translation initiation codon of exon 2 of the Prg4, proteoglycan 4 (megakaryocyte stimulating factor, articular superficial zone protein). The resulting chimeric animals were crossed to FLP recombinase expressing mice (ACT-FLPe) mice (on an unknown genetic background) to remove the Frt flanked PGK-neo selection cassette. |
When maintaining a live colony, these mice can be bred as homozygotes.
When using the STOCK Prg4tm1(GFP/cre/ERT2)Abl/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #022757 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Prg4<tm1(GFP/cre/ERT2)Abl> |
Frozen Mouse Embryo | STOCK Prg4<tm1(GFP/cre/ERT2)Abl>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Prg4<tm1(GFP/cre/ERT2)Abl>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Prg4<tm1(GFP/cre/ERT2)Abl>/J | $3373.50 |
Frozen Mouse Embryo | STOCK Prg4<tm1(GFP/cre/ERT2)Abl>/J | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.