基于转录组测序挖掘雌性鸡胚性腺不对称发育中的重要lncRNA及其靶基因  

Discovery of important lncRNA and their target genes related to the asymmetric gonadal development of female chicken embryos based on RNA-seq analysis

作　　者：韩紫燕 鲍海港[1] HAN Ziyan;BAO Haigang(College of Animal Science and Technology,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业大学动物科技学院,北京100193

出　　处：《黑龙江畜牧兽医》2024年第1期36-41,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家现代农业产业技术体系建设专项(CARS-40)。

摘　　要：为了挖掘雌性鸡胚性腺不对称发育中的重要长链非编码RNA(lncRNA)及其靶基因,试验以采集的胚胎孵化第9天(E9阶段)的雌性鸡胚两侧性腺为研究对象,采用转录组测序(RNA-seq)技术和生物信息学方法对雌性鸡胚左右两侧性腺进行转录组分析预测lncRNA,运用DESeq2 v1.30.1软件鉴定两侧性腺间差异表达的mRNA和lncRNA,预测差异表达lncRNA的顺式和反式靶基因,对靶基因进行GO功能注释和KEGG信号通路分析;通过实时荧光定量PCR扩增验证重要靶基因的表达。结果表明:转录组分析预测出653个基因间lncRNA,467个内含子lncRNA,1339个反义lncRNA。雌性鸡胚左右两侧性腺间有1856个差异表达mRNA和443个差异表达lncRNA。GRIA1、DIO3、FSHR、LHX9、CYP17A1和TOX3等229个基因为差异表达lncRNA的靶基因。两侧性腺差异表达lncRNA的靶基因主要富集到了类固醇代谢过程、细胞内雌激素受体信号通路的正向调节、代谢通路等过程。实时荧光定量PCR扩增验证重要靶基因的表达结果与RNA-seq结果基本一致。说明雌性鸡胚左右两侧性腺具有不同表达水平和特异性表达的mRNA和lncRNA,它们可能影响雌性鸡胚左右两侧性腺的不对称发育。In order to explore the important lncRNA and its target genes in the asymmetric development of female chicken embryo’s gonads,the gonads on both sides of female chicken embryo at E9 stage were taken as the research object in the experiment.The transcriptome sequencing(RNA-seq)technology and bioinformatics methods were used to analyze and predict the lncRNAs of the left and right gonads of female chicken embryos.And the DESeq2 v1.30.1 software was used to identify the differential expression mRNAs and lncRNAs between the gonads of both sides.The cis and trans target genes of differential expression lncRNAs were predicted.GO functional annotation and KEGG enrichment analysis on the target genes were performed.The expressions of important target genes were verified through quantitative real-time PCR(qRT-PCR).The results showed that 653 intergenic lncRNAs,467 intronic lncRNAs,and 1339 antisense lncRNAs were predicted by RNA-seq.There were 1856 differential expression mRNAs and 443 differential expression lncRNAs between the left and right gonads of female chicken embryos.229 genes including GRIA1,DIO3,FSHR,LHX9,CYP17A1,and TOX3 were found to be target genes for differential expression lncRNAs.The differential expression lncRNAs’target genes in both gonads were mainly enriched in steroid metabolism,positive regulation of intracellular estrogen receptor signaling pathways,and metabolic pathways.The expression of mRNAs confirmed by quantitative real-time PCR was basically consistent with the results of RNA-seq.These results indicated that the left and right gonads of female chicken embryos had different levels of expression and specific expression of mRNAs and lncRNAs,which might affect the asymmetric development of the left and right gonads of female chicken embryos.

关 键 词：转录组测序 鸡胚 雌性性腺 不对称发育 长链非编码RNA 

分 类 号：S831.2[农业科学—畜牧学]