猪lncRNA LOC102157897表达、定位及其对PEDV复制的调控作用OA北大核心CSTPCD
Expression,localization and its effect of porcine lncRNA LOC102157897 on PEDV replication
猪流行性腹泻(porcine epidemic diarrhea,PED)是由猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)引起的高度传染性疾病,前期基于组学测序筛选出1个关键lncRNA(LOC102157897),有关其表达定位以及对PEDV复制的调控作用目前尚不清楚.首先在组织和细胞水平上测定PEDV感染前后LOC102157897的表达水平;其次利用lncLocator软件预测和核质分离试验分析其细胞定位情况,并验证其表达水平与PEDV复制的关系;最后利用转录组测序筛选其下游靶基因及信号通路.结果表明:PEDV感染可极显著提高猪空肠组织和IPEC-J2细胞中LOC102157897的表达水平;LOC102157897主要定位于细胞核中,这可能与其生物学功能有关;成功构建了LOC102157897干扰细胞系,且干扰后IPEC-J2细胞中PEDV复制水平呈极显著下降.表达水平下调有利于提高宿主细胞对PEDV感染的抵抗能力.转录组测序分析显示,LOC102157897干扰前后存在151个差异表达基因,主要参与病毒蛋白互作、趋化因子信号、白细胞受体互作、白细胞介素17信号等免疫通路,并根据功能注释和差异程度,筛选出2个组蛋白基因H2AC18和H3C14.综上,这一研究确定了 1个PEDV抗性相关lncRNA分子LOC102157897,并初步探究了其功能和下游调控机制,为揭示lncRNA在PEDV感染宿主过程中的重要作用以及为今后制定PEDV的抗病育种工作策略和筛选分子标记物奠定基础.
Porcine epidemic diarrhea(PED)is a highly contagious disease caused by porcine epidemic diarrhea virus(PEDV),and one key lncRNA(LOC102157897)was previously selected based on omics sequencing,however its expres-sion localization as well as the regulation of PEDV replication are unknown.In this study,LOC102157897 expression levels were measured before and after PEDV infection at the tissue and cellular levels,followed by lncLocator software prediction and nucleocytoplasmic segregation analysis of its cellular localization,and then RNA interference was used to investigate the relation-ship between its expression levels and PEDV replication.The results showed that PEDV infection could significantly increase the expression level of LOC102157897 in porcine intestinal tissues and IPEC-J2 cells(P＜0.01).LOC102157897 was mainly local-ized in the nucleus,which may be related to its biological function;LOC102157897 interfering cell line was successfully constructed,and the PEDV replication level in IPEC-J2 cells showed a very significant decrease after interference.Down-regulation of expression levels was beneficial to improve host cell resistance to PEDV infection.Transcriptomic sequencing analysis showed that 151 differentially expressed genes existed before and after LOC102157897 interference,which were mainly involved in viral protein interaction,chemokine signaling,leukocyte receptor interaction,interleukin 17 signaling and oth-er immune pathways.Two histone genes,H2AC18 and H3C14,were screened according to functional annotation and degree of difference.Therefore,we preliminarily identified a PEDV resistance-associated lncRNA molecule-LOC102157897,and found that its down-regulated expression level was beneficial to provide host cell resistance to PEDV infection,and this study aimed to reveal the important role of lncRNAs in the host process of PEDV infection and lay the foundation for the devel-opment of disease resistance breeding strategies and screening molecular markers for PEDV in the future.
黄东璋;包文斌;吴正常
江苏农牧科技职业学院,江苏泰州 225300扬州大学动物科学与技术学院,江苏扬州 225009
畜牧业
猪流行性腹泻病毒长链非编码RNA表达调控
pigporcine epidemic diarrhea viruslong non-coding RNAexpressive regulation
《扬州大学学报（农业与生命科学版）》 2024 (005)
82-89 / 8
江苏省农业科技自主创新资金项目[CX(23)3084]
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