Construction and identification of synovial tissue conditional Grk2 knockout mice
- VernacularTitle:滑膜细胞条件性敲除Grk2基因小鼠的构建和鉴定
- Author:
Shu-jun ZUO
1
;
Wei-kang WANG
1
;
Jin-tao GU
1
;
Fu-yuan GUO
1
;
Hao-zhou GUO
1
;
Chen-chen HAN
1
;
Wei WEI
1
Author Information
- Publication Type:Journal Article
- Keywords: G protein-coupled receptor kinase 2; synovial tis-sue; synovial cells; Cre-loxP system; CRISPR/Cas9; condi-tioned knockout mice
- From: Chinese Pharmacological Bulletin 2025;41(6):1194-1199
- CountryChina
- Language:Chinese
- Abstract: Aim To construct and analyze the genotype of G protein-coupled receptor kinase 2(GRK2)conditional knockout mice in synoviocytes,and to provide an animal model for stud-ying the function of GRK2 in synoviocytes.Methods Grk2flox/+mice were bred to generate Grk2flox/flox mice,Grk2flox/flox mice were bred to Col1a1-iCre+mice,Grk2flox/+Col1a1-iCre+mice were bred to Grk2flox/flox mice.Grk2flox/flox Col1a1-iCre+mice were ob-tained as target mice.DNA was extracted and amplified by PCR to identify the genotype.Western blot was used to verify the effect of Grk2 knockout in synovium,liver and kidney tissues.HE staining was used to detect the effects of Grk2 conditional knockout in synovial cells on ankle synovium,liver and kidney tissues.Multiple immunofluorescence was used to detect GRK2 expression in synovial cells.Results The results of gene iden-tification showed that Grk2flox/flox Col1a1-iCre+mice had both Flox and Col1a1-iCre genotypes.Western blot results showed that GRK2 expression decreased in synovial tissues of Grk2flox/flox Col1a1-iCre+mice,but there was no significant change in the expression of GRK2 in liver and kidney tissues.HE staining showed that Grk2flox/flox Col1a1-iCre+mice had no significant pathological changes in the ankle synovium,liver and kidney.The results of multiple immunofluorescence showed that GRK2 expression in synovial cells of Grk2flox/flox Col1a1-iCre+mice de-creased.Conclusion Grk2 conditional knockout mice in syno-viocytes are successfully constructed and identified,which pro-vides an animal model for further study of the role of GRK2 in synovial-related diseases.
