STOCK Slc34a1^{tm1(EGFP/cre/ERT2)Bhum}/J

Stock number: 032285
Product name: SLC34a1^GCE
Research areas: Developmental Biology Research, Internal/Organ Research, Research Tools

Description

The SLC34a1^GCE mice express an eGFPCreERT2 fusion protein (EGFP and creERT2 fusion protein) from the Slc34a1 promoter/enhancer elements. These mice may be useful for studying fully differentiated proximal tubule epithelial cells and their role in repair after injury.

Detailed description

The SLC34a1^GCE mice express an eGFPCreERT2 fusion protein (EGFP and creERT2 fusion protein) from the Slc34a1 promoter/enhancer elements. Slc34a1 encodes a sodium-dependent inorganic phosphate transporter only expressed only in differentiated kidney epithelial stem cells in the proximal tubule. Heterozygous mice are viable and fertile. Homozygous mice have hypophosphatemia and the donating investigator has observed some spontaneous death in homozygotes. Cre-ER^T2 fusion gene activity is inducible, observed in the same cells only following tamoxifen administration. When SLC34a1^GCE mice are bred with mice containing loxP-flanked sequence, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Slc34a1-expressing cells of the offspring.

EGFP in this strain is detectable by immunohistochemistry, and is not detectable by direct fluorescence.

Of note, a similar strain with constitutively active Cre recombinase expression driven by the Slc34a1 promoter is available as Stock No. 040319.

Development

The SLC34a1^GCE targeting vector was designed to insert an eGFPCreER^T2 (GCE) fusion protein coding sequence and an frt-flanked PGKneobpA selection cassette into the initiation codon of the solute carrier family 34 (sodium phosphate), member 1 (Slc34a1) targeted gene. Specifically, this targeting vector contains an enhanced green fluorescent protein (EGFP) sequence and a CreER^T2 fusion gene (Cre-ER^T2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain). This construct was electroporated into F1 hybrid (C57BL/6J x 129Sv)-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with FLPo-expressing mice to remove the neo cassette. Resulting SLC34a1^GCE mice were bred to C57BL/6NJ mice (Stock No. 005304) females to remove the FLPo transgene and mice were further maintained on a mixed genetic background. Cryopreserved sperm was sent to The Jackson Laboratory Repository. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes.

Allele

Symbol: Slc34a1^{tm1(EGFP/cre/ERT2)Bhum}
Name: targeted mutation 1, Benjamin Humphreys
MGI accession ID: MGI:5544917
Generation method: Targeted
Attributes: Recombinase-expressing; Reporter; Inducible
Synonyms: SLC34a1^GCE
Marker symbol: Slc34a1
Marker name: solute carrier family 34 (sodium phosphate), member 1
Marker synonyms: FRTS2; HCINF2; Na/Pi cotransporter; NaPi-2; NaPi2A; NAPI-3; NaPi-IIa; NPHLOP1; NPT2; NPTIIa; renal Na+/Pi transporter; SLC11; Slc17a2
Chromosome: 13
Strain of origin: (C57BL/6J x 129/Sv)F1
Site of expression: Expressed only in differentiated kidney epithelial stem cells in the proximal tubule after tamoxifen administration.
Molecular note: An eGFP/cre/ERT2 (GCE) fusion gene with an Frt-flanked neo-polyA cassette were inserted into the ATG translation initiation codon in exon 2 of the Slc34a1 locus by homologous recombination in ES cells. FLPo-recombinase removed the neo cassette.