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
景新阳 1莫家远 2王楠 3刘永刚 4牟玉莲 5林鑫 6黄雷 7朱振东 8李奎7
作者信息
- 1. 青岛农业大学动物科技学院,青岛,266109;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
- 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;广西大学动物科学技术学院,南宁,530004
- 3. 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;中国农业科学院北京畜牧兽医研究所,北京,100193
- 4. 云南农业大学动物科技学院,昆明,650201
- 5. 中国农业科学院北京畜牧兽医研究所,北京,100193
- 6. 天津农学院动物科学与动物医学学院,天津,300392
- 7. 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
- 8. 青岛农业大学动物科技学院,青岛,266109
- 折叠
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M.stn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.
Abstract
Myostatin(Mstn),a negative regulator of skeletal muscle growth and development within the TGF-β superfamily member,plays a crucial role in modulating the development of skeletal muscles.Myostatin can impact muscle development,manifesting pheno-typically as traits such as double-muscling in individuals.To investigate the impact of Mstn gene knock-out on the muscle transcriptome levels,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.Further-more,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 Smtnl1 genes were significantly enriched in 27 muscle function related pathways.Dgat2 and Slc27a1 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 differentia-tion.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 Slc27a1.
关键词
小鼠(Mus/musculus)/C2C12细胞/Mstn基因/骨骼肌/脂质代谢Key words
Mice(Mus musculus)/C2C12 cell/Mstn gene/Skeletal muscle/Lipid metabolism引用本文复制引用
出版年
2024
NETL
NSTL
万方数据