The Wnt10bG2aCE knock-in allele was designed to both abolish Wnt10b gene expression and express EGFP and Cre-ERT2 proteins from the Wnt10b promoter in adult prostate and bladder.
Andrew P McMahon, University of Southern California
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing, Reporter, Inducible, Null/Knockout, RMCE-ready) | Wnt10b | wingless-type MMTV integration site family, member 10B |
The Wnt10bG2aCE 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-ERT2 proteins from the Wnt10b promoter/enhancer elements. WNT10B encodes a signaling protein involved in the regulation of cell fate and patterning during embryogenesis. Heterozygous Wnt10bG2aCE 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-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 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).
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-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 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)-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. This strain may be segregating for A from the ES cell used in it's
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | 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. |
Allele Name | targeted mutation 1, Andrew P McMahon |
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Allele Type | Targeted (Recombinase-expressing, Reporter, Inducible, Null/Knockout, RMCE-ready) |
Allele Synonym(s) | Wnt10btm1(GFP,cre/ERT2)Amc; Wnt10b-G2aCE |
Gene Symbol and Name | Wnt10b, wingless-type MMTV integration site family, member 10B |
Gene Synonym(s) | |
Promoter | Wnt10b, wingless-type MMTV integration site family, member 10B, mouse, laboratory |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | 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. |
Strain of Origin | C57BL/6N-Atm1Brd |
Chromosome | 15 |
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. |
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. |
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.
When using the Wnt10b-G2aCE mouse strain in a publication, please cite the originating article(s) and include JAX stock #028116 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Wnt10b<tm1(cre/ERT2)Amc> |
Frozen Mouse Embryo | C57BL/6-Wnt10b<tm1(cre/ERT2)Amc>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Wnt10b<tm1(cre/ERT2)Amc>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Wnt10b<tm1(cre/ERT2)Amc>/J | $3373.50 |
Frozen Mouse Embryo | C57BL/6-Wnt10b<tm1(cre/ERT2)Amc>/J | $3373.50 |
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