Method for constructing a partially humanized mouse model of LDLR gene
By performing gene fragment replacement and crispercas9 point mutation on the LDLR gene in C57 mice, a partially humanized mouse model of the LDLR gene was constructed, which solved the problem of mismatch in human and mouse models and achieved the need to study the pathogenic mechanism of point mutations in the body.
Patent Information
- Application Number
- CN202310199719.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-03-05
- Publication Date
- 2025-08-19
- Estimated Expiration
- 2043-03-05
AI Technical Summary
The prior art is difficult to construct a matching human and mouse model to study the pathogenic mechanism of point mutations in LDLR genes, especially due to the unconserved mutation sites between human and mouse, which leads to research difficulties.
By substituting gene fragments of exons 13 to 14 of the LDLR gene of C57 mice, and using crispercas9 to perform point mutations on the LDLR c.1988-8 locus, a partially humanized mouse model of LDLR gene was constructed.
A mouse model of partially humanized LDLR gene was successfully constructed, and some humanized groups experienced obvious hyperlipidemia, and the wild type did not change the blood lipid level, meeting the research needs.
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Abstract
Description
Technical Field
[0001] The present invention relates to a method for constructing a humanized animal model, and in particular to a method for constructing a partially humanized LDLR gene mouse model. Background Art
[0002] Research on the pathogenicity of LDLR gene point mutations is an important part of further understanding familial hypercholesterolemia. However, since the pathogenic sites in some clinical patients are relatively rare and prone to non-conservation between humans and mice, especially point mutations in introns of adjacent gene exons, the construction of partially humanized LDLR mice is an in vivo disease research model for such mutation sites. Summary of the Invention
[0003] The present invention provides a method for constructing a partially humanized LDLR gene mouse model.
[0004] A method for constructing a partially humanized LDLR gene mouse model, the method comprising:
[0005] LDLR c.1988-8T>A is a clear pathogenic site, but this mutation is not conserved between humans and mice, making it difficult to construct a matched mouse model to study the pathogenic mechanism of this point mutation in vivo. Therefore, the gene segment from exon 13 to exon 14 of the LDLR gene in C57 mice was replaced with the human gene segment from exon 13 to exon 14, and the LDLR c.1988-8 site was point mutated using CrisperCas9. Blood biochemistry results showed that some humanized point mutation groups showed significant hyperlipidemia, while some humanized wild-type groups did not change blood lipid levels.
[0006] Beneficial technical effects of the present invention:
[0007] Blood biochemistry results showed that some humanized point mutation groups showed obvious hyperlipidemia, while some humanized wild-type groups did not change blood lipid levels. BRIEF DESCRIPTION OF THE DRAWINGS
[0008] Figure 1 Comparison of the non-conserved LDLR c.1988-8T>A gene loci between human and mouse;
[0009] Figure 2 A scheme for partial humanized gene replacement of the mouse LDLR gene and construction of a partially humanized LDLRc.1988-8T>A point mutation;
[0010] Figure 3 Total cholesterol and triglyceride levels at baseline and after one month of high-fat diet feeding in mice, as well as serum gross profiles. Implementation Method
[0011] A method for constructing a partially humanized LDLR gene mouse model, the method comprising:
[0012] LDLR c.1988-8T>A is a clear pathogenic site, but this mutation is not conserved between humans and mice, making it difficult to construct a matched mouse model to study the pathogenic mechanism of this point mutation in vivo. Therefore, the gene segment from exon 13 to exon 14 of the LDLR gene in C57 mice was replaced with the human gene segment from exon 13 to exon 14, and the LDLR c.1988-8 site was point mutated using CrisperCas9. Blood biochemistry results showed that some humanized point mutation groups showed significant hyperlipidemia, while some humanized wild-type groups did not change blood lipid levels.
[0013] Comparison of the non-conserved LDLR c.1988-8T>A gene loci between human and mouse Figure 1 shown.
[0014] The scheme for partial humanization gene replacement of mouse LDLR gene and construction of partial humanized LDLR c.1988-8T>A point mutation are as follows Figure 2 shown.
[0015] The total cholesterol and triglyceride levels of mice at baseline and after one month of high-fat diet feeding, as well as the serum gross Figure 3 shown.
Claims
1. A method for constructing a partially humanized mouse model of the LDLR gene, the method being: LDLR c.1988-8T>A is a clear pathogenic site, but this mutation site is not conserved between humans and mice, making it impossible to construct a matching mouse model to study the pathogenic mechanism of the point mutation at the in vivo level. Therefore, the gene fragment from exon 13 to exon 14 of the C57 mouse LDLR gene was replaced with the human gene fragment from exon 13 to exon 14, and crispercas9 was used to perform a point mutation on the LDLR c.1988-8 site.
Citation Information
Patent Citations
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