This Phd1f/Phd2f/Phd3f triple floxed strain may be useful for generating conditional mutations in applications related to the study hypoxia inducible factor and oxygen homeostasis.
Guo-Hua Fong, Univ CT Health Center
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Egln1 | egl-9 family hypoxia-inducible factor 1 |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), No functional change) | Egln2 | egl-9 family hypoxia-inducible factor 2 |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), No functional change) | Egln3 | egl-9 family hypoxia-inducible factor 3 |
Prolyl hydroxylase domain (PHD) proteins catalyze oxygen-dependent prolyl hydroxylation
of hypoxia-inducible factor 1α and 2α subunits. Under normoxic conditions, hypoxia-inducible factor α subunits are targeted for degration by prolyl hydroxylation. The prolyl hydroxylase domain proteins encoded by Egln2, Egln1, and Egln3 have overlapping function in hydroxylating both HIF1A and HIF2A. These Phd1f/Phd2f/Phd3f mice carry floxed alleles for the 3 genes. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have the floxed sequences of the 3 genes deleted in the cre-expressing tissues. Mice that are homozygous for the floxed mutations are viable and fertile. For each of these mutations, removal of the floxed sequence creates a null allele.
This triple floxed mutant line was generated by breeding three independently created mutant lines, described below.
For the Egln2tm2Fong (Egln2floxN or Phd1f) allele, a targeting vector designed by Dr. Guo-Hua Fong (Univ CT Health Center) containing a FRT site flanked NEO cassette was utilized. A loxP site and this selection cassette was inserted downstream of exon 3 of the targeted gene, and another loxP site was inserted upstream of exon 3. This construct was electroporated into (129S6/SvEvTac x C57BL/6NCrl)F1 derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were used to generate chimeras by tetraploid aggregation technique. Germline mice were crossed with transgenic mice expressing Flpe under the control of beta-actin promoter (on an unknown genetic background) to remove the FRT site flanked NEO cassette. The mice were then crossed to CD-1 mice to breed out the FLPe recombinase transgene.
For the Egln1tm2Fong (Phd2f) allele, a targeting vector designed by Dr. Guo-Hua Fong (Univ CT Health Center) containing a FRT site flanked NEO cassette was utilized. A loxP site and this selection cassette was inserted downstream of exon 2 of the targeted gene, and another loxP site was inserted upstream of exon 2. This construct was electroporated into (129S6/SvEvTac x C57BL/6NCrl)F1 derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were used to generate chimeras by tetraploid aggregation technique. The mice were then crossed to transgenic mice
expressing Flpe recombinase under the control of beta-actin promoter (on an unknown genetic background) to remove the FRT site flanked NEO cassette. The mice were then crossed to CD-1 mice to breed out the FLPe recombinase transgene.
For the Egln3tm2Fong (Phd3f) allele, a targeting vector designed by Dr. Guo-Hua Fong (Univ CT Health Center) containing a FRT site flanked NEO cassette was utilized. A loxP site and this selection cassette was inserted downstream of exon 2 of the targeted gene, and another loxP site was inserted upstream of exon 2. This construct was electroporated into (129S6/SvEvTac x C57BL/6NCrl)F1 derived G4 embryonic stem (ES) cells which were transiently transfected with a FLPe recombinase vector to remove the NEO selection cassette. ES cells that had successfully undergone FLPe-mediated recombination and no longer retained the cassette were used to generate chimeras by tetraploid aggregation technique. The mice were then crossed to CD-1 mice.
Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
Allele Name | targeted mutation 2, Guo-Hua Fong |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | Egln1fl; Phd2f |
Gene Symbol and Name | Egln1, egl-9 family hypoxia-inducible factor 1 |
Gene Synonym(s) | |
Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Chromosome | 8 |
Molecular Note | An frt-flanked neo cassette with a 5' loxP site was inserted downstream of exon 2 and an additional loxP site was inserted upstream of exon 2. Subsequent germ-line, cre-mediated recombination resulted in the creation of Egln1tm1Fong. |
Allele Name | targeted mutation 2, Guo-Hua Fong |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | Egln2fl; Egln2floxN; Phd1fl |
Gene Symbol and Name | Egln2, egl-9 family hypoxia-inducible factor 2 |
Gene Synonym(s) | |
Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Chromosome | 7 |
Molecular Note | A loxP site was inserted upstream of exon 3. An additional loxP and a neo cassette were inserted downstream of exon 3. This allele was used to generate Egln2tm2Fong. |
Allele Name | targeted mutation 2, Guo-Hua Fong |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | Egln3fl; Egln3floxN; PHD3fl |
Gene Symbol and Name | Egln3, egl-9 family hypoxia-inducible factor 3 |
Gene Synonym(s) | |
Strain of Origin | (129S6/SvEvTac x C57BL/6NCrl)F1 |
Chromosome | 12 |
Molecular Note | A loxP site was inserted upstream of exon 2. An additional loxP site and a neo cassette were inserted downstream of exon 2. This allele was used to generate Egln3tm1Fong. |
When maintaining a live colony, these mice can be bred as homozygous for all 3 alleles.
When using the Phd1, Phd2, Phd3 Triple floxed mouse strain in a publication, please cite the originating article(s) and include JAX stock #028097 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Egln2<tm2Fong> , Egln1<tm2Fong> and Egln3<tm2Fong> |
Frozen Mouse Embryo | STOCK Egln2<tm2Fong> Egln1<tm2Fong> Egln3<tm2Fong>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Egln2<tm2Fong> Egln1<tm2Fong> Egln3<tm2Fong>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Egln2<tm2Fong> Egln1<tm2Fong> Egln3<tm2Fong>/J | $3373.50 |
Frozen Mouse Embryo | STOCK Egln2<tm2Fong> Egln1<tm2Fong> Egln3<tm2Fong>/J | $3373.50 |
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