摘要
长链非编码RNA(lncRNA,非编码RNA≥200 bp)以前被认为是转录中的“噪音”。然而,近年来已经发现了相当多的具有重要功能的lncRNA。LncRNA在疾病、发育、细胞分化和先天免疫反应中起着至关重要的作用。本研究通过使用TopHat2软件和Cufflinks软件分析草鱼的转录组数据,鉴定了与免疫相关的长链非编码RNA。筛选出一个lncRNA序列,命名为ISG152-L2,它位于草鱼ISG152的上游。使用综合基因组学查看器(IGV)测定ISG152-L2和草鱼ISG152的同步表达。研究了ISG152-L2的共表达,并确定了包括草鱼ISG152在内的相关基因网络,功能富集分析表明,这些基因在免疫相关过程中功能富集。用poly(I:C)刺激时,ISG152-L2在被测组织(脑,眼,肠,鳃,心脏,肝胰腺,脾脏和肾脏)中上调。同时,ISG152-L2和ISG152在CIK细胞受到poly(I:C)刺激后均上调。双荧光素酶基因检测发现,CIK细胞中的ISG152-L2显著增强了ISG152启动子活性,并且这种增强取决于ISG152-L2的特定620~1 190 bp区域。免疫共沉淀试验表明,草鱼ISG152可以在体外IRF3相互作用。此外,isg152-l2过表达增加了CIK细胞中IRF3的蛋白质水平,而ISG152-L2的干扰则产生了相反的结果。总之,我们筛选了一个免疫相关的lncRNA,即ISG152-L2,它位于ISG152的上游,并通过诱导其启动子的活性,上调IRF3的蛋白表达水平来增强ISG152的表达。
Long non-coding RNAs(lncRNAs;non-coding RNAs≥200 bp)were previously considered as transcriptional noise.However,a considerable number of lncRNAs with important functions have been identified in recent years.LncRNAs play vital roles in diseases,development,cell differentiation and innate immune responses,while the functional investigation of fish lncRNAs still remains limited.In this study,grass carp lncRNAs associated with immunity were identified by analyzing the transcriptome data of grass carp(Ctenopharyngodon idella)using TopHat2 and Cufflinks softwares.An lncRNA sequence was screened out,named ISG152-L2,located in the upstream of C.idella ISG152.The synchronized expression of ISG152-L2 and ISG152 was determined using the Integrative Genomics Viewer(IGV).The co-expression of ISG152-L2 was investigated,and the related genes network,including ISG152,was identified.Functional enrichment analysis indicated that these genes were functionally enriched in immunity-related processes.Upon stimulation with poly(I:C),ISG152-L2 was upregulated in the tested tissues(brain,eye,intestine,gill,heart,liver,spleen and kidney).Meanwhile,ISG152-L2 and ISG152 were both up-regulated following poly(I:C)stimulation in CIK cells.Dual-luciferase reporter assays found that ISG152 promoter activity was significantly enhanced by ISG152-L2 in CIK cells and this enhancement depended on the particular 620~1190 bp region of ISG152-L2.Co-immunoprecipitation assays indicated that ISG152 can interact with IRF3 in vitro.Furthermore,ISG152-L2 overexpression increased the protein level of IRF3 in CIK cells,whereas interference of ISG152-L2 had the opposite consequences.In conclusion,we screened an immune-related lncRNA ISG152-L2 which was located in the upstream of ISG152;further research found that ISG152-L2 could enhance ISG152 expression by inducing its promoter activity and up-regulating IRF3 protein level related to the body weight.Based on the results,it can be used as candidate loci for applying to molecular markers assisted breeding of g
作者
胡霁欢
胡美玲
史骁
周鹏程
常凯乐
向阳
徐小文
HU Jihuan;HU Meiling;SHI Xiao;ZHOU Pengcheng;CHANG Kaile;XIANG Yang;XU Xiaowen(School of Life Science,Nanchang University,Nanchang 330031,China)
出处
《南昌大学学报（理科版）》
CAS
北大核心
2023年第3期272-282,共11页
Journal of Nanchang University(Natural Science)
基金
国家自然科学基金地区资助项目(32160872)。
