This triple floxed mutant line was generated by breeding three independently created mutant lines, described below.



For the Egln2^tm2Fong (Egln2^floxN or Phd1^f) 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 Egln1^tm2Fong (Phd2^f) 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 Egln3^tm2Fong (Phd3^f) 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).

• None Available

Additional Information

• Considerations for Choosing Controls

• Clever D; Roychoudhuri R; Constantinides MG; Askenase MH; Sukumar M; Klebanoff CA; Eil RL; Hickman HD; Yu Z; Pan JH; Palmer DC; Phan AT; Goulding J; Gattinoni L; Goldrath AW; Belkaid Y; Restifo NP. 2016. Oxygen Sensing by T Cells Establishes an Immunologically Tolerant Metastatic Niche. Cell 166(5):1117-1131.e14PubMed: 27565342MGI: J:234541
• Duan LJ; Takeda K; Fong GH. 2014. Hematological, hepatic, and retinal phenotypes in mice deficient for prolyl hydroxylase domain proteins in the liver. Am J Pathol 184(4):1240-50PubMed: 24508125MGI: J:208042
• Takeda K; Ho VC; Takeda H; Duan LJ; Nagy A; Fong GH. 2006. Placental but Not Heart Defects Are Associated with Elevated Hypoxia-Inducible Factor {alpha} Levels in Mice Lacking Prolyl Hydroxylase Domain Protein 2. Mol Cell Biol 26(22):8336-46PubMed: 16966370MGI: J:114669