Construction of a myocardial and skeletal muscle-specific Cyr61 gene knockout mouse model
10.16352/j.issn.1001-6325.2025.06.0720
- VernacularTitle:心肌和骨骼肌特异性Cyr61基因敲除小鼠模型的构建
- Author:
Diyan XU
1
;
Wenli ZHANG
;
Yidan ZUO
;
Zhen SU
Author Information
1. 温州医科大学附属第一医院肾内科,浙江温州 325000
- Keywords:
Cyr6;
myocardium;
skeletal muscle;
gene conditional knockout mice
- From:
Basic & Clinical Medicine
2025;45(6):720-726
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a conditional knockout mice model of cysteine-rich angiogenesis inducer 61(Cyr61,also known as Ccn1)gene in myocardium and skeletal muscle regulated by CRE recombinant enzyme.Methods C57BL/6 mice were used to create CKmm-Cre+/-Cyr61flox/flox mice by using CRISPR/Cas9 technology.The successful construction of conditional knockout mice was confirmed by identifying the Cyr61flox/flox,3'LoxP and Cre enzyme sequences in mice.The knockout efficiency of Cyr61 was confirmed by Western blot.The skeletal mus-cle function of Cyr61 knockout mice was evaluated and following related indicators in the myocardium and skeletal muscle were detected by Western blot:Aging(p53,p21),inflammatory response(TNF-α,IL-18,IL-1β),fibrosis(vimentin,TGF-β,α-SMA,COL1).Results The mice were successfully bred and identified.In comparison with the control group,Cyr61 protein level showed a significant decrease in both myocardium and skeletal muscle in the experimental group(P＜0.01).Compared with the control group,the experimental group mice showed increased skeletal muscle grip strength(P＜0.05),enhanced maximum single contraction force(P＜0.01),and increased average cross-sectional area of the anterior tibialis muscle(P＜0.05).The expression level of p21,TNF-α,IL-18,IL-1β,TGF-β,and COL1 proteins in the myocardium and skeletal muscle of the experimental group was all lower than those of the control group(P＜0.05).Conclusions Myocardial and skeletal muscle-specific Cyr61 gene condi-tional knockout mice were successfully constructed based on Cre-LoxP technology and heritable trait could be passed stably.
