Basic Information
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Description
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- The exons 2-3 of mouse Prokr2 gene that encode signal peptide, extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hPROKR2 mice. The promoter and 5’UTR region of the mouse gene are retained. The human PROKR2 expression is driven by endogenous mouse Prokr2 promoter, while mouse Prokr2 gene transcription and translation will be disrupted.
- Mouse Prokr2 mRNA was detectable in wild-type C57BL/6JNidfc mice. Human PROKR2 mRNA was detectable in homozygous B-hPROKR2 mice.
- PROKR2 protein was detected in brain, olfactory bulb, lymph nodes, testis, spleen, colon of wild-type C57BL/6JNifdc mice and homozygous B-hPROKR2 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-hPROKR2 mice. The exons 2-3 of mouse Prokr2 gene that encode signal peptide, extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hPROKR2 mice. The promoter and 5’UTR region of the mouse gene are retained. The human PROKR2 expression is driven by endogenous mouse Prokr2 promoter, while mouse Prokr2 gene transcription and translation will be disrupted.
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mRNA expression analysis
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Strain specific analysis of PROKR2 mRNA expression in wild-type C57BL/6JNifdc mice and homozygous B-hPROKR2 mice by RT-PCR. Brain RNA was isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hPROKR2 mice (H/H). Mouse Prokr2 mRNA was only detectable in wild-type C57BL/6JNifdc mice, and human PROKR2 mRNA was exclusively detectable in homozygous B-hPROKR2 mice but not in wild-type C57BL/6JNifdc mice.
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Protein expression analysis
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Western blot analysis of PROKR2 protein expression in homozygous B-hPROKR2 mice. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hPROKR2 mice (H/H), and then analyzed by western blot with anti-PROKR2 antibody (Novus Biologicals, NBP1-45218). PROKR2 was detected in brain, olfactory bulb, lymph nodes, testis, spleen, colon of wild-type C57BL/6JNifdc mice and homozygous B-hPROKR2 mice.
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Protein expression analysis in neutrophils
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CD300E expression analysis in F1 B-hCD300E mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+) and F1 B-hCD300E mice, and analyzed by flow cytometry with anti-CD300E antibody (BD Biosciences, 566315). hCD300E was not detectable in neutrophils.
