Basic Information
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Description
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- PLVAP is a single-pass type II membrane homodimer glycoprotein (50-55 kDa) primarily expressed on most endothelial cells, which is endothelial cell-specific membrane protein involved in the formation of the diaphragms that bridge endothelial fenestrae.
- The exons 1-5 of mouse Plvap gene that encode the extracellular domain were replaced by human counterparts in B-hPLVAP mice including human 3’UTR region. The genomic region of mouse Plvap gene that encodes transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were retained. The human PLVAP expression is driven by endogenous mouse Plvap promoter, while mouse Plvap gene transcription and translation will be disrupted.
- Human PLVAP protein was exclusively detectable in the heart, liver, spleen, lung, and kidney of homozygous B-hPLVAP mice.
- Mouse Plvap mRNA was only detectable in wild-type C57BL/6JNifdc mice. Human PLVAP mRNA was only detectable in homozygous B-hPLVAP mice.
- 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-hPLVAP mice. The exons 1-5 of mouse Plvap gene that encode the extracellular domain were replaced by human counterparts in B-hPLVAP mice including human 3’UTR region. The genomic region of mouse Plvap gene that encodes transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were retained. The human PLVAP expression is driven by endogenous mouse Plvap promoter, while mouse Plvap gene transcription and translation will be disrupted.
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Protein expression analysis
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Strain specific PLVAP expression analysis in homozygous B-hPLVAP mice by western blot. Heart, liver, spleen, lung, and kidney were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hPLVAP mice (H/H), and analyzed by western blot with anti-PLVAP antibody(CST, 38238). 40 μg total proteins were loaded for western blotting analysis. hPLVAP and deglycosylated hPLVAP were exclusively detectable in these tissues of homozygous B-hPLVAP mice but not in wild-type C57BL/6JNifdc mice.
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mRNA expression analysis
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Strain specific analysis of PLVAP mRNA expression in wild-type mice and B-hPLVAP mice by RT-PCR. Lung RNA was isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hPLVAP mice (H/H). Mouse Plvap mRNA was only detectable in wild-type C57BL/6JNifdc mice, and human PLVAP mRNA was exclusively detectable in homozygous B-hPLVAP mice but not in wild-type C57BL/6JNifdc mice.
