Mstn基因敲除小鼠肌肉和C2C12细胞的转录特征分析  

Analysis of Transcription Characteristics of Muscle and C2C12 Cells in Mstn Gene Knock-out Mice

作　　者：景新阳 莫家远 王楠 刘永刚[5] 牟玉莲 林鑫[6] 黄雷 朱振东 李奎 JING Xinyang;Mo Jiayuan;WANG Nan;LIU Yonggang;MU Yulian;LIN Xin;HUANG Lei;ZHU Zhendong;LI Kui(College of Animal Science and Technology,Qingdao Agricultural University,Qingdao,266109;Shenzhen Branch,Guangdong Laboratory of Lingnan Modern Agriculture,Key Laboratory of Livestock and Poultry Multi-omics of Ministry of Agriculture and Rural Affairs,Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen,518124;College of Animal Science and Technology,Guangxi University,Nanning,530004;Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing,100193;College of Animal Science and Technology,Yunnan Agricultural University,Kunming,650201;College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin,300392)

机构地区：[1]青岛农业大学动物科技学院,青岛266109 [2]岭南现代农业科学与技术广东省实验室深圳分中心,农业农村部畜禽生物组学重点实验室,中国农业科学院(深圳)农业基因组研究所,深圳518124 [3]广西大学动物科学技术学院,南宁530004 [4]中国农业科学院北京畜牧兽医研究所,北京100193 [5]云南农业大学动物科技学院,昆明650201 [6]天津农学院动物科学与动物医学学院,天津300392

出　　处：《基因组学与应用生物学》2024年第11期1779-1793,共15页Genomics and Applied Biology

基　　金：国家自然科学基金项目(32130102);深圳市杰出人才培养项目;国家重点研发计划项目(2021YFD1301200)共同资助。

摘　　要：肌肉生长抑制素(Myostatin,Mstn)作为TGF-β超家族的成员,是一种调控骨骼肌生长发育的负调节因子,其失活可以促进肌肉的发育,在个体上表现为双肌臀等表型。为了探究Mstn基因敲除对肌肉细胞和组织转录组的影响,本研究利用CRISPR/Cas9技术构建了3个Mstn基因敲除的C2C12细胞株,并采集了12只Mstn基因敲除小鼠(Mus musculus)(6公6母)的腓肠肌,分别进行转录组测序分析。结果显示,Mstn基因敲除C2C12细胞增殖能力显著高于野生型细胞的。在Mstn基因敲除的C2C12细胞、公鼠和母鼠中分别鉴定到329、240、265个差异表达基因(differentially expressed genes,DEGs)。Mstn基因敲除公鼠和母鼠中鉴定到50个共有DEGs,在下调表达基因中,Myh7、Lmod2、Barx2、Myh2、Myl3和Smtnl1基因显著富集到27条肌肉功能相关通路;Dgat2和Slc27a1显著富集到8条脂质代谢相关通路。Mstn基因敲除C2C12细胞和小鼠中鉴定到4个共有DEGs,其中Fam83d基因是成肌细胞增殖、分化的关键调控基因。综上所述,Mstn基因敲除既可以通过下调Myh7、Lmod2和Fam83d等基因参与肌肉发育调控,也能够通过下调Dgat2和Slc27a1基因调控脂质代谢。Myostatin(Mstn),a negative regulator of skeletal muscle growth and development within the TGF-βsuperfamily menber,plays a crucial role in modulating the development of skeletal muscles.Myostatin can impact muscle development,manifesting phenolevels,this study applied CRISPR/Cas9 technology in generating three strains of C2C12 cell with Mstn gene knock-out.Additionally,the gastrocnemius muscles from 12 Mstn gene knock-out mice(6 males,6 females)were collected for transcriptome sequencing.The results showed that the proliferative capacity of Mstn gene knock-out cells was significantly higher than that of wild-type cells.Furthermore,transcriptome sequencing revealed that 329,240,265 differentially expressed genes(DEGs)were found in the cells,male mice,and female mice,respectively.50 DEGs were both identified in Mstn gene knock-out male and female mice.Among the down regulated genes,Myh7,Lmod2,Barx2,Myh2,Myl3,and Smtnll genes were significantly enriched in 27 muscle function related pathways.Dgat2 and Slc27al were significantly enriched in 8 lipid metabolism related pathways.In addition,4 DEGs were identified in Mstn gene knock-out cells and mice,among which the Fam83d gene was a key regulatory gene for myoblast proliferation and differentiation.Above all.Mstn gene knock-out can participate in muscle development regulation by down regulated genes such as Myh7,Lmod2,and Fam83d,as well as regulating lipid metabolism by down regulated genes Dgat2 and Slc27al.

关 键 词：小鼠(Mus musculus) C2C12细胞 MSTN基因 骨骼肌 脂质代谢 

分 类 号：Q78[生物学—分子生物学]