Basic Information
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Description
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- PTH1R is a receptor for both PTH and PTHrP, and PTH binding promotes Ca2+ release from bone, mediated by osteoclast formation, which is a predicted 66 – 68 kDa member of the G-protein coupled receptor family.
- The exons 4-16 of mouse Pth1r gene that encode extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hPTH1R mice. The promoter, 5’UTR region and signal peptide of the mouse gene are retained. The human PTH1R expression is driven by endogenous mouse Pth1r promoter, while mouse Pth1r gene transcription and translation will be disrupted.
- Mouse Pth1r mRNA was detectable in wild-type C57BL/6N mice. Human PTH1R mRNA was detectable in homozygous B-hPTH1R mice.
- PTH1R protein was detected in kidney and lung of wild-type C57BL/6N mice and homozygous B-hPTH1R mice, as the antibody can cross-recognize both human and mouse PTH1R.
- Application: This product is used for pharmacodynamics and safety evaluation of drugs.
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Targeting strategy
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Gene targeting strategy for B-hPTH1R mice. The exons 4-16 of mouse Pth1r gene that encode extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hPTH1R mice. The promoter, 5’UTR region and signal peptide of the mouse gene are retained. The human PTH1R expression is driven by endogenous mouse Pth1r promoter, while mouse Pth1r gene transcription and translation will be disrupted.
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mRNA expression analysis in humanized B-hPTH1R mice
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Species specific analysis of PTH1R gene expression in wild-type C57BL/6N mice and homozygous humanized B-hPTH1R mice by RT-PCR. Kidney was collected from wild-type C57BL/6N mice (+/+) and homozygous B-hPTH1R mice (H/H). Mouse PTH1R mRNA was detectable in wild-type C57BL/6N mice. Human PTH1R mRNA was detectable only in homozygous B-hPTH1R mice, but not in wild-type C57BL/6N mice.
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Protein expression analysis
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Strain specific PTH1R expression analysis in homozygous humanized B-hPTH1R mice by western blot. Liver, kidney, spleen and lung were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hPTH1Rmice (H/H) and analyzed by western blot with anti-PTH1Rantibody(Abcam, ab189924). m/hPTH1R was detectable in kidney and lung of wild-type C57BL/6N mice and homozygous B-hPTH1R mice.
Strain specific PTH1R expression analysis in homozygous humanized B-hPTH1R mice by western blot. Kidney and lung were collected from wild-type C57BL/6N mice (+/+), and heterozygous B-hPTH1R mice (H/+) and homozygous B-hPTH1R mice (H/H) and analyzed by western blot with anti-PTH1R antibody(Abcam, ab189924). m/hPTH1R was detectable in kidney and lung of wild-type C57BL/6N mice, heterozygous B-hPTH1R mice and homozygous B-hPTH1R mice.