C57BL/6-Wnt10b^{tm1(cre/ERT2)Amc}/J

Stock number: 028116
Product name: Wnt10b-G2aCE
Research areas: Developmental Biology Research, Internal/Organ Research, Research Tools

Description

The Wnt10b^G2aCE knock-in allele was designed to both abolish Wnt10b gene expression and express EGFP and Cre-ER^T2 proteins from the Wnt10b promoter in adult prostate and bladder.

Detailed description

The Wnt10b^G2aCE knock-in allele was designed to both abolish wingless-type MMTV integration site family, member 10B (Wnt10b) gene expression and express individual EGFP and Cre-ER^T2 proteins from the Wnt10b promoter/enhancer elements. WNT10B encodes a signaling protein involved in the regulation of cell fate and patterning during embryogenesis. Heterozygous Wnt10b^G2aCE mice are viable and fertile. The donating investigator has not attempted to make this strain homozygous. Weak EGFP fluorescence is seen in the skin of 5 day old mice. Cre-ER^T2 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 prostate, epididymis, bladder and skin. Specifically, when tamoxifen is induced at P3, Cre-mediated recombination occurs in skin and keratin-8 (Krt8)-expressing cells of the prostate starting at P5, and in the epididymis at 12 weeks of age.

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

Development

An exchange vector was designed to be inserted downstream of exon 1 of the wingless-type MMTV integration site family, member 10B (Wnt10b) 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-ER^T2 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-A^tm1Brd-derived JM8A1.N3 knockout first reporter embryonic stem (ES) cells obtained from the Knockout Mouse Project (KOMP) consortium containing clone EPD0314_2_A09. Recombined ES cells, with the neo cassette removed and resistant to puro, were injected into B6(Cg)-Tyr^c-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. This strain may be segregating for A from the ES cell used in it's

Allele

Symbol: Wnt10b^{tm1(cre/ERT2)Amc}
Name: targeted mutation 1, Andrew P McMahon
MGI accession ID: MGI:5660815
Generation method: Targeted
Attributes: Recombinase-expressing; Reporter; Inducible; Null/Knockout; RMCE-ready
Marker symbol: Wnt10b
Marker name: wingless-type MMTV integration site family, member 10B
Chromosome: 15
Strain of origin: C57BL/6N-A^tm1Brd
Expressed genes: GFP,Green Fluorescent Protein,; cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: Cre-ERT2 fusion protein and EGFP are expressed from endogenous Wnt10b promoter/enhancer elements in the prostate, epididymis, bladder and skin.
Molecular note: An exchange vector was designed to be inserted downstream of exon 1 of the gene using dual-recombinase mediated cassette exchange (dRMCE). Specifically, the exchange vector contained (from 5 to 3) an FRT site, a viral F2A oligopeptide (to mediate ribosomal skipping), an enhanced green fluorescent protein 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 cassette and a loxP site were inserted at the 3' end of the exchange vector.
General note: The 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-derived JM8A1.N3 knockout first reporter embryonic stem (ES) cells obtained from the Knockout Mouse Project (KOMP) consortium containing clone EPD0314_2_A09. Recombined ES cells lack the neo cassette and are resistant to puromycin.