宿主环形RNA circCAMSAP2对猪丁型冠状病毒增殖的影响  

Effects of host circular RNA circCAMSAP2on proliferation of porcine deltacoronavirus

作　　者：刁素静 杜柳阳 万涛[1,2] 周继勇[1,2] 顾金燕 DIAO Sujing;DU Liuyang;WAN Tao;ZHOU Jiyong;GU Jinyan(Animal Medicine Center,Zhejiang University,Hangzhou 310058,China;MOA Key Laboratory of Animal Virology,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学动物医学中心,浙江杭州310058 [2]浙江大学农业农村部动物病毒学重点实验室,浙江杭州310058

出　　处：《中国兽医学报》2024年第12期2526-2533,共8页Chinese Journal of Veterinary Science

基　　金：“十四五”国家重点研发计划资助项目(2021YFD1801102);国家自然科学基金资助项目(32272971)。

摘　　要：旨在探究宿主环形RNA(circular RNA,circRNA)对猪丁型冠状病毒(porcine deltacoronavirus,PDCoV)增殖的影响。从前期获得的PDCoV感染ST细胞的circRNA差异表达谱中筛选得到1个在PDCoV感染后表达水平下调的circRNA circCAMSAP2进行研究。利用Ensembl数据库在线分析和Sanger测序对circCAMSAP2的基因来源及接头序列进行验证;利用核质分离试验、RNase R水解试验对circCAMSAP2的分子特征进行鉴定;利用RT-qPCR对circCAMSAP2在PDCoV感染前后的差异表达水平进行验证;利用circCAMSAP2的过表达及敲低试验研究circ-CAMSAP2对PDCoV复制的影响。结果显示,circCAMSAP2是由CAMSAP2基因2号外显子经反向剪接而形成的circRNA分子,长度为260nt,具有RNase R耐受性,主要定位于细胞质中。PDCoV感染抑制circCAMSAP2的表达。过表达circCAMSAP2可以抑制PDCoV的增殖,沉默circCAMSAP2可以促进PDCoV的增殖。结果表明,猪源circCAMSAP2负调控PDCoV的增殖,是设计抗PDCoV药物的潜在新靶点。The purpose of this study is to investigate the impact of host circular RNA(circRNA)on the proliferation of porcine deltacoronavirus(PDCoV).Among the significantly downregulated circRNAs identified during PDCoV infection from circRNA expression profiles in ST cells,circCAMSAP2was selected for further analysis.The gene origin and junction sequence of circCAMSAP2 were validated using the online Ensembl database and Sanger sequencing.The molecular characteristics of circCAMSAP2were determined through nucleoplasmic separation assays and RNase R tests.Additionally,the differential expression of circCAMSAP2during PDCoV infection was confirmed using RT-qPCR.Overexpression and knockdown experiments were conducted to investigate the regulatory role of circCAMSAP2in PDCoV replication.The results indicated that circCAMSAP2is a circRNA derived from exon 2of the CAMSAP2gene through reverse splicing.It is resistant to RNase R,has a length of 260nucleotides,and is predominantly located in the cytoplasm.circCAMSAP2expression is downregulated during PDCoV replication.Overexpression of circ-CAMSAP2inhibited PDCoV proliferation,while knockdown promoted it in ST cells.These findings suggest that porcine circCAMSAP2acts as a negative regulator of PDCoV replication and could serve as a potential target for the development of anti-PDCoV therapeutics.

关 键 词：环形RNA circCAMSAP2 猪丁型冠状病毒 病毒增殖 

分 类 号：S852.65[农业科学—基础兽医学]