Basic Information
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Description
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- Ketohexokinase (KHK) is the first enzyme in fructose metabolism, primarily found in the liver, kidneys, and small intestine. It phosphorylates fructose to fructose-1-phosphate (F1P), a substrate for fatty acid synthesis linked to non-alcoholic fatty liver disease (NAFLD). Excess liver fat is associated with obesity, insulin resistance, and type 2 diabetes.
- Gene editing strategy: The exons 1-8 of mouse Khk gene that encode the whole molecule (ATG to STOP codon), including 3’UTR were replaced by human counterparts in B-hKHK mice. The promoter and 5’UTR region of the mouse gene were replaced by human counterparts.
- KHK was detectable in liver, kidney, small intestine and liver of wild-type C57BL/6JNifdc mice and homozygous B-hKHK mice, but not in homozygous B-Khk KO mice.
- Human KHK targeted nucleic acid drugs (provided by the client) were efficacious in B-hKHK mice.
- Application: This product is used for pharmacodynamics and safety evaluation of type 2 diabetes, obesity and other metabolic diseases.
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Targeting strategy
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Gene targeting strategy for B-hKHK mice. The exons 1-8 of mouse Khk gene that encode the whole molecule (ATG to STOP codon), including 3’UTR were replaced by human counterparts in B-hKHK mice. The promoter and 5’UTR region of the mouse gene were replaced by human counterparts. The human KHK expression is driven by human KHK promoter, while mouse Khk gene transcription and translation will be disrupted.
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Protein expression analysis
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Strain specific KHK expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hKHK mice by western blot. Liver, kidney and small intestine were collected from wild-type C57BL/6JNifdc mice (+/+), homozygous B-hKHK mice (H/H) and homozygous B-Khk KO mice (-/-), and analyzed by western blot with anti-KHK antibody (abcam, ab197593). KHK was detectable in liver, kidney, small intestine and liver of wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hKHK mice (H/H), but not in homozygous B-Khk KO mice (-/-).
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mRNA expression analysis
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Species specific analysis of KHK gene expression in wild-type C57BL/6JNifdc mice and homozygous humanized B-hKHK mice by RT-PCR. Liver was collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hKHK mice (H/H). Mouse Khk mRNA was detectable only in wild-type C57BL/6JNifdc mice. Human KHK mRNA was detectable only in homozygous B-hKHK mice, but not in wild-type C57BL/6JNifdc mice.
