Basic Information
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Description
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- BMP9 is a secreted protein and after binding to its receptors, SMAD family transcription factors were recruited and activated, leading to gene expression.
- Homozygous mice expressed the human BMP9 and developed normally.
- This model is useful in studying the role of BMP9 in liver fibrosis.
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Targeting strategy
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Gene targeting strategy for B-hBMP9 mice. The exons 1-2 of mouse Bmp9 gene that encode the whole molecule (ATG to STOP codon), including 3’UTR were replaced by human counterparts in B-hBMP9 mice. The promoter and 5’UTR region of the mouse gene are retained. The human BMP9 expression is driven by endogenous mouse Bmp9 promoter, while mouse Bmp9 gene transcription and translation will be disrupted.
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mRNA expression analysis
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Strain specific analysis of BMP9 mRNA expression in wild-type C57BL/6N mice and homozygous B-hBMP9 mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hBMP9 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human BMP9 primers. Mouse Bmp9 mRNA was detectable only in wild-type C57BL/6N mice. Human BMP9 mRNA was detectable only in homozygous B-hBMP9 mice but not in wild-type mice.
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Protein expression analysis
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Western blot analysis of BMP9 protein expression in homozygous B-hBMP9 mice. Plasma and serum were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hBMP9 mice (H/H), (male, 7-week-old, n=2) and then analyzed by western blot with anti-BMP9 antibody (Abcam, ab207318). 20 μg total proteins were loaded for western blotting analysis. The antibody used showed cross-recognition for mouse and human BMP9.
