甲型流感病毒A/WSN/1933(H1N1)感染A549细胞的circRNA表达谱分析与鉴定  

Expression profiling analysis and characterization of circRNA in A549 cells infected by influenza virus strain A/WSN/1933(H1N1)

作　　者：张晓婷 郑一青 曾亦然 常丽凤 陈纳 卢渊录 罗皓炜 高紫荷 平继辉[1] ZHANG Xiaoting;ZHENG Yiqing;ZENG Yiran;CHANG Lifeng;CHEN Na;LU Yuanlu;LUO Haowei;GAO Zihe;PING Jihui(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《畜牧与兽医》2024年第7期102-111,共10页Animal Husbandry & Veterinary Medicine

基　　金：“十四五”国家重点研发计划(2021YFD1800205)。

摘　　要：环状RNA(circRNA)已被证明在众多生物过程中起着重要的调控作用,而circRNA在甲型流感病毒(IAV)复制过程中的功能尚不清楚。本研究旨在利用高通量测序分析IAV感染A549细胞后的circRNA表达谱变化来挖掘能够影响病毒复制的circRNAs并对其进行初步功能鉴定。利用测序结果,对全部circRNAs进行特征分析、差异表达筛选;对差异circRNAs进行GO分析、KEGG分析;对候选circRNAs进行qPCR、RT-PCR验证及Sanger测序鉴定,并通过空斑试验确定影响病毒复制的circRNAs。结果:本试验共筛选到11 630条差异表达circRNAs,对其中6条circRNA进行了鉴定,最终确定novel_circ_007428具有抗病毒作用。本研究对甲型流感病毒感染A549细胞后的circRNA表达谱进行了分析与鉴定,为进一步研究circRNA在流感病毒感染中发挥的调控作用奠定了基础。Circular RNA(circRNA)has been shown to play an important role in the regulation of various biological processes.However,the function of circRNA in influenza A virus(IAV)replication is unknown and remains to be investigated.Therefore,the aim of this study was to utilize high-throughput sequencing to analyze the changes in circRNA expression profiles in A549 cells infected by IAV to find the circRNAs that can affect the viral replication.All circRNAs obtained by RNA sequencing were characterized,and were screened for differentially expressed circRNAs.The differently expressed circRNAs were analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).Candidate circRNAs were selected for qPCR,RT-PCR validation and Sanger sequencing validation,and circRNAs affecting viral replication were identified by knockdown experiments.The results showed that a total of 11630 candidate circRNAs were screened out;six of the circRNAs were experimentally confirmed to be correctly formed into circular RNAs;and novel_circ_007428 was confirmed to have antiviral effects.In conclusion,this study analyzed and characterized the circRNA expression profile in A549 cells infected by IAV,which provided a basis for further research on the molecular mechanisms of circRNAs in influenza virus infection.

关 键 词：环状RNA 甲型流感病毒 基因芯片测序 A549 表达谱 novel_circ_007428 

分 类 号：S852.65[农业科学—基础兽医学]