B6;DBA-Tg(Tmem100-EGFP/cre/ERT2)30Amc/J

Stock number: 014159
Research areas: Research Tools

Description

This Tmem100-GCE transgenic strain expresses GFP and tamoxifen inducible Cre recombinase in the developing kidney and reproductive tracts and may be useful in studies of genitourinary development.

Detailed description

These transgenic mice express the eGFPCreER^T2 (Enhanced Green Fluorescent Protein and cre/ESR1) fusion gene under the direction of the mouse Tmem100, transmembrane protein 100, promoter. Transgene expression is detected in developing nephrons of the kidney, Wolfian and Mullerian ducts and vasculature of the urogenital system of hemizygous 15.5 dpc embryos. GFP fluorescence is detected in embryos 15.5dpc in age in the proximal region of the early renal vesicle, at later stages eGFP is present in the parietal epithelium adjacent to Wilms Tumor (WT1) positive cells demarcating the future podocytes of Bowman's capsule and is limited to the Tmem100 expression domain. Tamoxifen inducible Cre recombinase activity is detected in the developing kidney (nephrogenic zone), vasculature and Wolfian or Mullerian duct of mutant embryos aged 15.5dpc. Possible cre recombinase activity is detected in small renal arteries. Other possible sites of expression have not been characterized. Mice hemizygous for the transgenic insert are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.



This strain was transferred from the collection of the GenitoUrinary Development Molecular Anatomy Project (GUDMAP).

Development

A targeting construct containing the eGFPCreER^T2 (GCE) cassette and FRT site flanked Kanamycin/neomycin selection cassette was inserted into the consensus start ATG codon of the gene in a BAC (RP23-183L17). The PGK-Neo/Kan cassette was removed by transient Flpe-recombinase expression. The resulting BAC transgene was microinjected into (C57BL/6 x DBA) F1donor eggs. Founder line 30 was subsequently established. The mice were then crossed to C57BL/6 for two generations. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Tg(Tmem100-EGFP/cre/ERT2)30Amc
Name: transgene insertion 30, Andrew P McMahon
MGI accession ID: MGI:4849986
Generation method: Transgenic
Attributes: Recombinase-expressing; Reporter; Inducible
Marker symbol: Tg(Tmem100-EGFP/cre/ERT2)30Amc
Marker name: transgene insertion 30, Andrew P McMahon
Chromosome: UN
Strain of origin: (C57BL/6 x DBA)F1
Expressed genes: cre,cre recombinase,bacteriophage P1; ESR1,estrogen receptor 1,human; GFP,Green Fluorescent Protein,
Site of expression: GFP and tamoxifen inducible cre recombinase is expressed in the developing kidney and reproductive tracts.
Molecular note: A targeting construct containing the eGFPCreERT2 (GCE) cassette and Frt site flanked Kanamycin/neomycin selection cassette was inserted into the consensus start ATG codon of the gene in a BAC (RP23-183L17). The PGK-Neo/Kan cassette was removed by transient Flpe-recombinase expression. The resulting BAC transgene was microinjected into (C57BL/6 x DBA)F1 donor eggs. Founder line 30 was subsequently established. Native GFP expression is detectable in the urogenital system of E15.5 embryos. Transgenic cre expression is detected in developing nephrons of the kidney, Wolfian and Mullerian ducts, and vasculature of the urogenital system of hemizygous 15.5 dpc embryos. Possible cre recombinase activity is detected in small renal arteries.