Basic Information
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Gene Targeting Strategy
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The human GPC3 CDS was inserted after exon 3 of the mouse Gpc3 gene in B-hGPC3 mice. This insertion disrupts the endogenous murine Gpc3 gene.
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mRNA Expression Analysis
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Species–specific GPC3 expression analysis in wild-type and humanized B-hGPC3 mice. Kidneys were isolated from wild-type and homozygous B-hGPC3 mice and analyzed by RT-PCR for GPC3 mRNA expression. Human GPC3 mRNA was exclusively detected in B-hGPC3 mice compared to wild-type mice.
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IHC analysis of GPC3 expression
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Immunohistochemical (IHC) analysis of GPC3 protein expression in C57BL/6 wild-type (WT) mice and B-hGPC3 mice. The liver and fetal liver were collected from C57BL/6 (+/+) and homozygous B-hGPC3 mice (H/H) and analyzed by IHC with anti-GPC3 antibody. Mouse GPC3 was detectable in WT mice, and human GPC3 was detectable in homozygous B-hGPC3 mice. The arrow indicates tissue cells with positive GPC3 staining (brown).
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Protein expression profile of GPC3
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Immunohistochemical (IHC) analysis of GPC3 protein expression in C57BL/6 wild-type (WT) mice and B-hGPC3 mice. Nine major tissues were collected from C57BL/6 mice and homozygous B-hGPC3 mice (female/male, 8-week-old), and analyzed by IHC with anti-GPC3 antibody. Humanized GPC3 was detected in the fetal liver (H) of homozygous B-hGPC3 mice (Figure 2). There was no detectable expression of hGPC3 in various tissues of C57BL/6 (Figure 1). A summary of the expression levels in various tissues of C57BL/6 and B-hGPC3 is presented in Table 1. Note: In both C57BL/6 and B-hGPC3 mice, staining can be observed in the heart. however, since the positive tissue (fetal liver) specifically recognizes hGPC3 with the antibody, it can be inferred that the staining in the heart is non-specific.
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Summary
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- mRNA expression analysis:
Mouse GPC3 mRNA was detectable in kidney of WT mice (+/+). Human GPC3 mRNA was exclusively detectable in homozygous B-hGPC3 mice (H/H) mice but not in WT mice (+/+).
- Protein expression analysis:
GPC3 was exclusively detectable in kidney, heart and brain from homozygous B-hGPC3 mice (H/H) and WT mice (+/+) due to the cross-reactivity of antibodies.
