MSTN基因编辑牛肌肉转录组测序及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2021.07.001

Abstract

Abstract: To explore the specific regulation mechanism of myostatin (MSTN) gene on muscle development of bovine,a total of ten healthy bovine were selected from the same farm,including five MSTN^-/- gene-edited bovine and five wild-type bovine,and extracted the RNA of bovine muscle tissue sample.Collecting the calf-hip muscle samples of two groups,and the transcriptome analysis were sequenced by Illumina high-throughput sequencing technology.Bioinformatics method was used to compare the differentially expressed genes between two groups,and Go and KEGG enrichment analysis were performed.Real-time quantitative PCR was used to verify the transcriptome sequencing data.A total of 18 071 genes were obtained from the gene-edited bovine and wild-type bovine.Under the conditions of log₂|FoldChange|≥ 1.48,a total of 406 significantly differentially expressed genes were screened,in which 347 genes were up-regulated and 59 genes were down-regulated.GO functional analysis results showed that total 915 functional categories were effected after MSTN gene editing (P<0.05).The differentially expressed genes were mainly involved in the binding,regulation of biological process,immune system process and other related functions.KEGG functional enrichment analysis results indicated that differential genes were involved in total of 211 pathways,mainly concentrated in cell adhesion molecule,chemokine,cytokine interaction and other signaling pathways.The differentially expressed genes might be related to cell growth and muscle development were further screened out,including CD14,KIT,CSF1R,FBP1,DUSP4,ULBP21,PRKCB,SPN,CHAD and SRC genes.Real-time quantitative PCR results showed that the expression levels of the differentially expressed genes were basically consistent with the transcriptome expression,indicating the reliability of sequencing results.The results of this study indicated that MSTN gene could mediate the expression of multiple downstream genes after exert its role,effecting the relevant signaling pathways and biological processes.The screened differentially expressed genes in this study could be used as candidate targets for further study of the regulation mechanism of skeletal muscle.

Key words: myostatin; transcriptome sequencing; gene screening; Real-time quantitative PCR

CLC Number:

MIAO Manning, GUO Yiwen, HU Debao, ZENG Yuhan, LI Xin, ZHANG Linlin, DING Xiangbin, GUO Hong. Transcriptome Sequencing and Bioinformatics Analysis of MSTN Gene-edited Bovine Muscles[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2271-2281.

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Transcriptome Sequencing and Bioinformatics Analysis of MSTN Gene-edited Bovine Muscles

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