心肌锚定重复序列蛋白基因心脏特异敲除小鼠模型构建与鉴定

目的 构建心肌锚定重复序列蛋白(CARP)基因心脏特异敲除小鼠模型,为研究CARP基因与心脏疾病的关系奠定基础.方法 构建CARP基因条件打靶载体pL253-CARP-LoxP电转入小鼠胚胎干细胞(ES)细胞,G418和GANC药物筛选阳性细胞克隆,Southern blot确定CARP基在条件打靶载体转染成功.胚胎注...

Construction and identification of mice model with cardiac disruption of cardiac ankyrin repeat protein

Abstract: Objective To establish the mice model with cardiac disruption of cardiac ankyrin repeat protein (CARP) and to provide reliable tools for studying the function of CARP gene in heart related diseases. Methods Construct the conditional target plasmid of PL253-CARP-LoxP and transfect to 129 embryonic stem cells. Positive clones selected by antibiotics of G418 and GANC were determined by Southern blot to confirm conditional disruption of CARP gene, and then were injected to the blastula. Chimeras were got and crossed with α-MHC-Cre transgenic mice, which expressed recombinase specifically in heart, and then the mice with cardiac deficiency of CARP gene (CARP-KO) will get. Results Six chimeras were got, and CARP-KO mice were got after crossing with α-MHC-Cre mice. CARP gene was disrupted in the heart of CARP-KO mice by semi-quantitative PCR and western blot. These mice displayed normal in embryonic development, body growth, and heart development. However the expression of hypertrophic markers of β-MHC and ANF increased significantly in CARP-KO mice. Conclusion: The mice model with specific disruption of CARP gene was generated successfully. The cardiac disruption of CARP gene did not interfere the development and growth of mice significantly, either for the heart development.