B6(Cg)-Egln1^{tm4.1(EGLN1*)Fsl}/Mmjax

Stock number: 040415
Product name: Phd2^Tib
Research areas: Cardiovascular Research, Cell Biology Research, Hematological Research

Description

Phd2^Tib knock-in (KI) mice have endogenous exon 1 of the Egln1 gene replaced with human exon 1 sequence that carries Tibetan D4E and C127S amino acid substitutions. This mouse line may be useful when studying oxygen-sensing and the hypoxia-inducible factor (HIF) pathway.

Of note, the donating investigator has also made a Phd2^WT mouse line available as RRID:MMRRC_075765-JAX, to serve as a control to this Phd2^Tib mouse line.

Detailed description

Prolyl hydroxylase domain protein 2 (PHD2), coded by Egln1, is a protein that plays a crucial role in regulating oxygen levels in the body by hydroxylating hypoxia-inducible factor alpha (HIF-α), which is a subunit of the hypoxia-inducible factor (HIF) complex. HIF is necessary for oxygen homeostasis by regulating metabolic adaptation, erythropoiesis (red blood cell production), respiration, and angiogenesis. These Phd2^Tib mice have a humanized exon 1, carrying a D4E/C127S double amino acid substitution found in high-altitude Tibetans, replacing endogenous exon 1 of the Egln1 gene.

Homozygous mice are viable and fertile. Phd2^Tib homozygotes display augmented hypoxic ventilatory response. These mice may be useful when studying oxygen-sensing and the hypoxia-inducible factor (HIF) pathway.

Of note, the donating investigator has also made a Phd2^WT mouse line available as RRID:MMRRC_075765-JAX, to serve as a control to this Phd2^Tib mouse line.

Development

The Phd2^Tib targeting vector was designed by replacing the endogenous exon 1 of the egl-9 family hypoxia-inducible factor 1 (Egln1) gene, with human exon 1 sequence that harbors a D4E/C127S double amino acid substitution. The targeting construct also contains a loxP-flanked neomycin selection cassette. The construct was electroporated into C57BL/6-derived EAP6 embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. Chimeric mice were bred to B6(Cg)-Tyr^c-2J/J mice to confirm germline transmission. Resulting Phd2^Tib mice were then crossed to the C57BL/6-Gt(ROSA)26Sor^{tm16(Cre)Arte} mouse line to remove the floxed neomycin cassette. The Cre recombinase allele was removed through selective breeding to C57BL/6J mice. The Phd2^Tib mouse line was maintained on a C57BL/6 background by the donating laboratory. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Egln1^{tm4.1(EGLN1*)Fsl}
Name: targeted mutation 4.1,Frank Lee
MGI accession ID: MGI:8186458
Generation method: Targeted
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: Phd2^Tib
Marker symbol: Egln1
Marker name: egl-9 family hypoxia-inducible factor 1
Marker synonyms: C1orf12; ECYT3; HALAH; Hif-p4h-2; HIFPH2; HIF-PH2; HPH2; HPH-2; ORF13; PHD2; PHD-2; SM20; SM-20; ZMYND6
Chromosome: 8
Strain of origin: C57BL/6
Molecular note: The targeting vector was designed to replace the endogenous exon 1 of the egl-9 family hypoxia-inducible factor 1 (Egln1) gene with a human exon 1 sequence that harbors a D4E/C127S double amino acid substitution. The targeting construct also consisted of a loxP-flanked neomycin selection cassette. The the floxed neomycin cassette was removed by Cre recombinase.