Construction of hyperuricemic mouse model with Uox gene knockout based on CRISPR/Cas9 system
10.3969/j.issn.1005-4847.2025.03.009
- VernacularTitle:基于CRISPR/Cas9系统构建Uox基因敲除的高尿酸血症小鼠模型
- Author:
Yiwei ZHANG
1
;
Weihu LONG
;
Donghong TANG
;
Shengtao FAN
;
Peng WANG
;
Chenyun WANG
;
Zheli LI
;
Zhangqiong HUANG
;
Yousong YE
Author Information
1. 北京协和医学院中国医学科学院医学生物学研究所,昆明 650118
- Publication Type:Journal Article
- Keywords:
hyperuricemia;
animal model;
CRISPR/Cas9;
gene knockout;
uricase
- From:
Acta Laboratorium Animalis Scientia Sinica
2025;33(3):411-419
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a uricase-deficient mouse model with stable inheritance using the CRISPR/Cas9 system,and evaluate its ability to simulate the disease characteristics of patients with hyperuricemia.Methods Double single guide RNAs(sgRNAs)were designed on both sides of exon 2～4 of the Uox gene.sgRNA and Cas9 mRNA for gene knockout were microinjected into the fertilized eggs of mice.After culture for 2～4 h,the embryos were transferred to surrogate mother mice to produce an F0 generation.Uox-knockout mice were identified by polymerase chain reaction and sequencing analysis.Positive mice were then mated with wild-type(WT)mice to produce an F1 generation,and heterozygous female and male F1 mice were then selected to obtain homozygous F2 mice.Serum and urine levels of uric acid,creatinine,and urea,and serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels were detected and compared between homozygous and wild-type mice.Pathological changes in kidney and liver tissues were observed by hematoxylin and eosin and Masson staining.Results Urine levels of serum uric acid(male:(4116.8±1928.1)μmol/L,P＜0.001;female:(2998.0±547.7)μmol/L,P＜0.01)and serum levels of uric acid(male:(478.4±114.6)μmol/L,P＜0.001;female:(507.7±129.6)μmol/L,P＜0.001),creatinine((91.8±55.6)μmol/L,P＜0.001),urea((28.6±13.9)mmol/L,P＜0.05),ALT((53.3±23.3)U/L,P＜0.01),and AST((203.3±70.3)U/L,P＜0.001)were significantly increased in Uox-/-mice compared with WT mice.Histopathological examination showed moderate hepatocyte degeneration in the liver,moderate-to-severe tubular cystic dilation,degeneration,and fibrosis in the kidney,glomerular hypertrophy and hyperplasia,small-vessel dilation and congestion,and infiltration of stromal monocytes and lymphocytes in Uox-/-mice.Conclusions We successfully established a homozygous uricase-deficient mouse strain using CRISPR/Cas9 technology,as a suitable animal model for research in the field of hyperuricemia.
