Mice hemizygous for the BAC Hsd17b1-icreERT2 transgene are viable, fertile, and normal in size with iCre-ERT2 expression directed by the hydroxysteroid (17-beta) dehydrogenase 1 (Hsd17b1) promoter/enhancer regions within the BAC transgene. These mice may be useful for studying gene function in ovarian granulose cells.
Emilio Casanova, Ludwig Boltzmann Institute for Cancer Re
Genetic Background | Generation |
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Allele Type |
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Transgenic (Recombinase-expressing, Inducible) |
Mice hemizygous for the BAC Hsd17b1-icreERT2 transgene are viable, fertile, and normal in size with iCre-ERT2 expression directed by the hydroxysteroid (17-beta) dehydrogenase 1 (Hsd17b1) promoter/enhancer regions within the BAC transgene. These BAC mice express 8 copies of iCre-ERT2 protein in granulose cells of the ovarian follicles. iCre-ERT2 fusion gene activity is inducible and is observed only following tamoxifen administration. When these mice are bred with mice containing loxP-flanked sequence, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the BAC-expressing cells of the offspring. These mice may be useful for studying gene function in ovarian granulose cells.
The iCre-ERT2 fusion protein consists of a codon-improved Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, iCre-ERT2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered.
Bacterial artificial chromosome (BAC) library (CHORI) was used to obtain a 200 kb BAC (#RP24-178B1) containing the entire mouse hydroxysteroid (17-beta) dehydrogenase 1 (Hsd17b1) gene. This BAC was altered to insert an icreERT2 fusion gene (iCre-ERT2; a codon improved Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain) and a polyadenylation signal (polyA) downstream of the initiation codon of Hsd17b1. This modified BAC was microinjected into fertilized (C57BL/6xCBA) F1 oocytes. Offspring were bred to C57BL/6NTac and founder line 3, containing 8 copies of the Hsd17b1- iCreERT2 BAC, was established. These mice were subsequently bred with C56BL/6NTac inbred mice for at least 6 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation.
Expressed Gene | ESR1, estrogen receptor 1, human |
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Expressed Gene | cre, cre recombinase, bacteriophage P1 |
Site of Expression | When mice carrying this transgene are bred with mice containing loxP-flanked sequence, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in granulose cells of the ovarian follicles. |
Allele Name | transgene insertion 3, Emilio Casanova |
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Allele Type | Transgenic (Recombinase-expressing, Inducible) |
Allele Synonym(s) | Hsd17b1-iCreERT2 |
Gene Symbol and Name | Tg(Hsd17b1-icre/ERT2)3Casa, transgene insertion 3, Emilio Casanova |
Gene Synonym(s) | |
Promoter | Hsd17b1, hydroxysteroid (17-beta) dehydrogenase 1, mouse, laboratory |
Expressed Gene | ESR1, estrogen receptor 1, human |
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
Site of Expression | When mice carrying this transgene are bred with mice containing loxP-flanked sequence, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in granulose cells of the ovarian follicles. |
Strain of Origin | (C57BL/6 x CBA)F1 |
Chromosome | UN |
Molecular Note | A 200kb BAC containing the Hsd17b1 gene was modified by homologous recombination to generate the construct. The open reading frame sequence for codon-improved cre recombinase (icre) fused to a mutated ligand binding domain (LBD) of the human estrogen receptor (ERT2) was inserted into the exon containing the transcription initiation codon of the Hsd17b1 gene. The initial targeting vector also contained the bovine growth hormone polyA signal and an Frt-flanked ampicillin cassette. The amp marker was subsequently removed from targeted by clones upon electroporation of a plasmid expressing Flp recombinase. Purified BAC clones were injected into (C57BL/6 x CBA)F1 oocytes. Three lines expressing the transgene were generated. Lines 1 and 2 were shown to contain 2 copies of the insert by Southern blot, while line 3 had 8 copies. Line 3 mRNA levels of icre/ERT2 were highest in ovaries and this line was used for subsequent analyses. |
Mutations Made By | Emilio Casanova, Ludwig Boltzmann Institute for Cancer Re |
When maintaining a live colony, hemizygous mice may be bred to wildtype (non-carrier) mice or C57BL/6NJ inbred mice (Stock No. 005304).
When using the B6N.Cg-Tg(Hsd17b1-icre/ERT2)3Casa/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #017310 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemizygous or Non carrier for Tg(Hsd17b1-icre/ERT2)3Casa |
Frozen Mouse Embryo | B6N.Cg-Tg(Hsd17b1-icre/ERT2)3Casa/J | $2595.00 |
Frozen Mouse Embryo | B6N.Cg-Tg(Hsd17b1-icre/ERT2)3Casa/J | $2595.00 |
Frozen Mouse Embryo | B6N.Cg-Tg(Hsd17b1-icre/ERT2)3Casa/J | $3373.50 |
Frozen Mouse Embryo | B6N.Cg-Tg(Hsd17b1-icre/ERT2)3Casa/J | $3373.50 |
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