利用CRISPRi干扰研究MyoG和Myf6基因对牛骨骼肌卫星细胞分化的影响  

Influence of MyoG and Myf6 Genes on the Differentiation of Bovine Skeletal Muscle Satellite Cell Using CRISPRi

作　　者：李冬晓[1] 李树峰[1] 佟慧丽[1] 张伟伟[1] 刘丹[1] 严云勤[1] 

机构地区：[1]东北农业大学生命科学学院细胞生物学实验室,哈尔滨150030

出　　处：《中国细胞生物学学报》2016年第3期292-302,共11页Chinese Journal of Cell Biology

基　　金：国家转基因专项"高产优质转基因肉牛新品种培育"(批准号:2014ZX08007-002)资助的课题~~

摘　　要：CRISPR干扰(clustered regularly interspaced short palindromic repeat interference,CRISPRi)技术因高效的基因干扰效率而成为基因功能研究的重要工具。Myo G、Myf6基因是生肌调节因子家族(myogenic regulatory factors,MRFs)的重要成员,是骨骼肌分化所必需的调控因子。该研究以牛骨骼肌卫星细胞为实验材料,探讨Myo G和Myf6基因在骨骼肌卫星细胞分化过程中的相互关系。构建Myo G、Myf6基因CRISPRi载体,分别转染牛骨骼肌卫星细胞,诱导其分化,Real-time PCR检测肌肉分化重要功能基因Myo G、Myf6、MYH2、Myo D的表达情况。结果表明,在牛骨骼肌卫星细胞分化期间,抑制Myo G基因表达将诱导Myf6基因的代偿性升高,但并不能完全弥补Myo G基因的缺少对肌肉分化产生的影响,而抑制Myf6基因表达则不会引起Myo G基因表达升高,这为肌肉分化机制的阐明提供了理论依据。CRISPRi（clustered regularly interspaced short palindromic repeat interference）, has become an important tool for the study of gene functions due to its efficiency. Myo G and Myf6 are important members of the myogenic regulatory factors family, which are necessary regulating factors for skeletal muscle differentiation. In this study, we set primary bovine skeletal muscle satellite cells as the experimental material to fully investigatethe interrelation between Myo G and Myf6. The CRISPRi vector of Myo G and Myf6 are contructed and transfected into bovine skeletal muscle satellite cells respectively, and induce differentiation, then detect the expression of Myo G, Myf6, MYH2, Myo D via Real-time PCR. The result shows that during differentiation of bovine skeletal muscle satellite cells, the inhibition of Myo G induces the compensatory increase of Myf6, though this can not offset the influence of Myo G on muscle differentiation. In contrast, the inhibition of Myf6 can not increase the expression ofMyo G. These results provide theoretical basis for the mechanism study of muscle differentiation.

关 键 词：MYOG Myf6 CRISPRi 牛 骨骼肌卫星细胞 分化 

分 类 号：S823[农业科学—畜牧学]