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Construction method and application of URAT1 humanized mouse model

A construction method and humanization technology, applied in chemical instruments and methods, biochemical equipment and methods, other methods of inserting foreign genetic materials, etc., can solve problems such as screening and evaluation

Pending Publication Date: 2022-03-22
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI +1
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Problems solved by technology

[0008] The main purpose of the present invention is to provide a method for constructing a URAT1 humanized mouse model, by replacing the human URAT1 coding gene with the corresponding mouse gene, and preparing a humanized mouse model of the specific uric acid transport receptor URAT1 , with human functional genes, can be used to screen and evaluate uric acid metabolism drugs targeting human URAT1, which solves the problem that rodents cannot be screened and evaluated on mice due to the difference in species properties between rodents and humans Issues with URAT1 Inhibitors in Humans

Method used

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  • Construction method and application of URAT1 humanized mouse model
  • Construction method and application of URAT1 humanized mouse model
  • Construction method and application of URAT1 humanized mouse model

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Embodiment

[0139] Reagents, Materials and Instruments

[0140] Animal Tissue / Cell Genomic DNA Extraction Kit (D1700 Beijing Solaibao Technology Co., Ltd.); 2×Rapid Taq Master Mix (P222 Nanjing Novizan Biotechnology Co., Ltd.); Uric Acid Detection Kit (C012-2 Nanjing Jiancheng Biological Technology Co., Ltd.); Engineering Research Institute); anti-human URAT1 antibody (200760-T08 Yiqiao Shenzhou). The synthesis and sequencing of all primers for the microscope slide scanner (3DHISTECH KFT) were completed by Qingke Biotechnology Co., Ltd.

[0141] Specific implementation steps

[0142] 1. Determination of the replacement region of the human URAT1 gene and the inserted human sequence

[0143] According to the function of human URAT1, the human gene URAT1 (SLC22A12 CDS, transcript NM_144585.4, located on human chromosome 11) was selected to replace the mouse URAT1 (NM_009203.3, located on mouse chromosome 19) gene, and the selected human The full-length amino acid sequence (Aa: 1-553; SEQ ...

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Abstract

The invention discloses a construction method of a URAT1 humanized animal model. The method comprises the following steps: (1) replacing a URAT1 gene on a background animal cell with an adult URAT1 gene; and (2) expressing the human URAT1 gene in the background animal cell and generating a humanized URAT1 protein, and reducing or eliminating the expression of an endogenous URAT1 gene in the background animal cell at the same time, thereby obtaining the URAT1 humanized animal model. The urate specific transporter URAT1 humanized mouse model constructed by the method has human functional genes, and can be used for URAT1 targeted drug screening, or curative effect evaluation of URAT1 targeted drugs and other drug combination, and subsequent pharmacological and toxicological research of drugs.

Description

technical field [0001] The present invention relates to the technical field of animal genetic engineering and genetic modification, in particular to a construction method based on a URAT1 humanized animal model and its application in the field of medicine. Background technique [0002] Gout is a group of diseases caused by purine metabolism disorders. Its clinical features are hyperuricemia and the resulting gouty acute arthritis recurrent attacks, tophi deposition, tophi chronic arthritis and joint deformities, often involving Kidney, causing chronic interstitial nephritis and uric acid nephrolithiasis. [0003] As a metabolic disease, the incidence of gout and hyperuricemia is increasing year by year in my country, which is a major factor affecting people's health. Specific inhibitors of uric acid transporter URAT1 (human uratetransporter 1) and organic anion transporter OTA (organic anion transporter) distributed on renal proximal tubule epithelial cells can reduce the r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/89C12N15/90C12N15/113C12N9/22A01K67/027A61K49/00
CPCC12N15/8509C12N15/89C12N15/907C12N15/113C12N9/22C07K14/47A01K67/0278A61K49/0008A01K2207/15A01K2217/072A01K2267/0393A01K2227/105C12N2310/20
Inventor 陈阿鹏德格晶
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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