The Spp2G2aCE knock-in allele was designed to both abolish Spp2 gene expression and express EGFP and Cre-ERT2 proteins from the Spp2 promoter in proximal tubule cells of the mouse kidney.
Andrew P McMahon, University of Southern California
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Recombinase-expressing, Reporter, Inducible, Null/Knockout, RMCE-ready) | Spp2 | secreted phosphoprotein 2 |
The Spp2G2aCE knock-in allele was designed to both abolish secreted phosphoprotein 2 (Spp2) gene expression and express individual EGFP and Cre-ERT2 proteins from the Spp2 promoter/enhancer elements. SPP2 encodes a serum phosphoprotein synthesized by the liver which is involved in bone growth, repair, and turnover. Heterozygous Spp2G2aCE mice are viable and fertile. The donating investigator has not attempted to make this strain homozygous, and mice may still be segregating for the Atm1Brd allele. Weak EGFP fluorescence is seen in proximal tubule cells of the mouse kidney and in liver cells at 19.5 days post coitum. Cre-ERT2 gene activity is inducible and can be observed following tamoxifen administration. As such, when these mice are bred with mice containing loxP-flanked sequences, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in proximal tubule cells in the embryonic and adult kidney.
This strain was transferred from the collection of the GenitoUrinary Development Molecular Anatomy Project (GUDMAP).
An exchange vector was designed to be inserted downstream of exon 1 of the secreted phosphoprotein 2 (Spp2) gene using dual-recombinase mediated cassette exchange (dRMCE). Specifically, the exchange vector contained (from 5’ to 3’) a frt site, a viral F2A oligopeptide (to mediate ribosomal skipping), an enhanced green fluorescent protein (EGFP) sequence, a viral T2A oligopeptide, and a Cre-ERT2 sequence (Cre recombinase fused to a G525R mutant form of the mouse estrogen receptor ligand binding domain). A rox-flanked puromycin resistance (puro) cassette and a loxP site were inserted at the 3' end of the exchange vector.
This exchange vector (along with the pDIRE plasmid containing an icre-expression cassette and a Flpo-expression cassette to facilitate insertion of the exchange vector) was electroporated into C57BL/6N-Atm1Brd-derived JM8A3.N1 knockout first reporter embryonic stem (ES) cells obtained from the European Conditional Mouse Mutagenesis (EUCOMM) consortium containing clone EPDO558_1_A04. Recombined ES cells, with the neo removed and resistant to puro, were injected into B6(Cg)-Tyrc-2J/J blastocysts and resulting chimeric mice were bred to C57BL/6J to establish a colony. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony. Mice may still be segregating for the Atm1Brd allele.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | When mice carrying this allele are bred with mice containing loxP-flanked sequences, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in proximal tubule cells in the embryonic and adult kidney. |
Allele Name | targeted mutation 1, Andrew P McMahon |
---|---|
Allele Type | Targeted (Recombinase-expressing, Reporter, Inducible, Null/Knockout, RMCE-ready) |
Allele Synonym(s) | Spp2G2aCE; Spp2tm1(GFP,cre/ERT2)Amc |
Gene Symbol and Name | Spp2, secreted phosphoprotein 2 |
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 | When mice carrying this allele are bred with mice containing loxP-flanked sequences, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in proximal tubule cells in the embryonic and adult kidney. |
Strain of Origin | C57BL/6N-Atm1Brd |
Chromosome | 1 |
General Note | This exchange vector (along with the pDIRE plasmid containing an icre-expression cassette and a Flpo-expression cassette to facilitate insertion of the exchange vector) was electroporated into C57BL/6N-Atm1Brd-derived JM8A3.N1 knockout first reporter embryonic stem (ES) cells obtained from the European Conditional Mouse Mutagenesis (EUCOMM) consortium containing clone EPDO558_1_A04). Recombined ES cells, with the neo removed and resistant to puro, were injected into B6(Cg)-Tyrc-2J/J blastocysts and resulting chimeric mice were bred to C57BL/6J to establish a colony. Recombined ES cells, with the neo removed and resistant to puro, were injected into B6(Cg)-Tyrc-2J/J blastocysts and resulting chimeric mice were bred toC57BL/6J to establish a colony. |
Molecular Note | The allele was generated by a dual recombinase-mediated cassette exchange protocol. The exchange vector contained (from 5' to 3') an frt site, a viral F2A oligopeptide (to mediate ribosomal skipping), an enhanced green fluorescent protein (EGFP) sequence, a viral T2A oligopeptide, and a cre/ERT2 sequence. A rox-flanked puromycin resistance (puro) cassette and a loxP site were inserted at the 3' end of the exchange vector. |
When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). The donating investigator has not attempted to make this strain homozygous. Mice may still be segregating for the Atm1Brd allele.
When using the C57BL/6-Spp2tm1(cre/ERT2)Amc/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #025209 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or Wildtype for Spp2<tm1(GFP,cre/ERT2)Amc> |
Frozen Mouse Embryo | C57BL/6-Spp2<tm1(cre/ERT2)Amc>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Spp2<tm1(cre/ERT2)Amc>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Spp2<tm1(cre/ERT2)Amc>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | C57BL/6-Spp2<tm1(cre/ERT2)Amc>/J Frozen Embryo | $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.