Construction of hepatocyte-specific Sirt3 gene knockout mouse model
10.19405/j.cnki.issn1000-1492.2024.03.003
- VernacularTitle:肝细胞特异性Sirt3基因敲除小鼠模型的构建
- Author:
Yaping XU
1
;
Yuhan WANG
;
Tingting CHEN
;
Nan LI
;
Pingping GAO
;
Ling LI
;
Hua WANG
;
Wuyi SUN
Author Information
1. 安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,合肥 230032
- Keywords:
hepatocyte;
Sirt3;
gene knockout;
Sirt3 Δhep mice;
Cre-loxP;
identification of genotype
- From:
Acta Universitatis Medicinalis Anhui
2024;59(3):384-390
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct hepatocyte-specific silence information regulator 3(Sirt3)gene knockout(Sirt3 Δhep)mice by Cre-loxP technique,and to provide an important animal model for further studying the biological function of the hepatocyte Sirt3 gene in diseases.Methods LoxP-labeled Sirt3flox/flox mice were mated with Alb-Cre homozygous(Alb-Cre+/+)mice,and the F1 generation Sirt3flox/-/Alb-Cre+/-mice were then mated with Sirt3flox/flox mice,and the F2 genotype of Sirt3flox/flox/Alb-Cre+/-mice were the Sirt3 Δhep mice constructed in this ex-periment.Sirt3flox/flox/Alb-Cre-/-(Sirt3flox/flox)mice were the control mice.Mouse tail genome DNA was extracted and PCR was used to identify the genotypes of the offspring mice.Immunofluorescence was used to detect Sirt3 ex-pression in mouse hepatocytes.Primary hepatocytes and tissue proteins of Sirt3 Δhep mice were extracted,and the ex-pression of Sirt3 in mouse hepatocytes and other tissues was verified by Western blot.HE staining was used to ob-serve mice's liver,heart,spleen,and lung tissue structure.Results Sirt3 Δhep mice were successfully identified.Immunofluorescence and Western blot results demonstrated a significant decrease in the expression of Sirt3 in the hepatocytes of these mice compared to the control group(P<0.01).At the same time,there was no significant difference in the expression of Sirt3 in the heart,spleen,kidney,and lung tissues of Sirt3 Δhep mice compared with the control group(P>0.05).The results of HE staining showed that the histological characteristics of the liver,heart,spleen,lungs,kidneys,and other major organs of Sirt3 Δhep mice were not significantly different from those of the control group mice.Conclusion Hepatocyte-specific Sirt3 gene knockout mice are successfully constructed,which provides an animal model to explore further the role and molecular mechanism of the hepatocyte Sirt3 gene in diseases.
