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| 1 | Expression Profile and Function Analysis of Long Non-coding RNAs in the Infection of Coxsackievirus B3显示文摘The roles of IncRNAs in the infection of enteroviruses have been barely demonstrated.In this study,we used coxsackievirus B3(CVB3),a typical enterovirus,as a model to investigate the expression profiles and functional roles of IncRNAs in enterovirus infection.We profiled IncRNAs and mRNA expression in CVB3-infected HeLa cells by IncRNA-mRNA integrated microarrays.As a result,700 differentially expressed IncRNAs(431 up-regulated and 269 down-regulated)and 665 differentially expressed mRNAs(299 up-regulated and 366 down-regulated)were identified in CVB3 infection.Then we performed IncRNA-mRNA integrated pathway analysis to identify potential functional impacts of the differentially expressed mRNAs,in which IncRNA-mRNA correlation network was built.According to IncRNA-mRNA correlation,we found that XLOC-001188,an IncRNA down-regulated in CVB3 infection,was negatively correlated with NFAT5 mRNA,an anti-CVB3 gene reported previously.This interaction was supported by qPCR detection following siRNA-mediated knockdown of XLOC-001188,which showed an increase of NFAT5 mRNA and a reduction of CVB3 genomic RNA.In addition,we observed that four most significantly altered IncRNAs,SNHG11,RP11-145F16.2,RP11-1023L17.1 and RP11-1021N1.2 share several common correlated genes critical for CVB3 infection,such as BRE and IRF2BP1.In all,our studies reveal the alteration of IncRNA expression in CVB3 infection and its potential influence on CVB3 replication,providing useful information for future studies of enterovirus infection. | Lei Tong Ye Qiu Hui Wang Yunyue Qu Yuanbo Zhao Lexun Lin Yan Wang Weizhen Xu Wenran Zhao Hongyan He Guangze Zhao Mary HZhang Decheng Yang Xingyi Ge Zhaohua Zhong | 2019 | Virologica Sinica2019，34，6： | 2 |
| 2 | 靶向3D基因shRNA慢病毒载体的构建及功能鉴定显示文摘本研究旨在筛选抑制B组柯萨奇病毒(coxsackievirus B,CVB)3D聚合酶表达的短发卡RNA(short hairpin RNA,shRNA)并构建其慢病毒表达载体,以抑制CVB的3D聚合酶表达。设计并合成3对针对3D基因的RNA干扰序列及相应的对照组序列,通过定量反转录-聚合酶链反应(quantitative reverse transcription-polymerase chain reaction,RT-qPCR)和蛋白免疫印迹法筛选出干扰效率最高的一对,合成shRNA序列。退火后,应用基因重组技术构建pLVTHM-3DshRNA重组质粒,经酶切鉴定及DNA测序后,将重组正确的pLVTHM-3DshRNA与psPAX2、pMD2.G共转染293T细胞,包装慢病毒并鉴定其对CVB3 3D基因表达的干扰效果。测序结果显示,成功构建了pLVTHM-3DshRNA重组慢病毒载体,并收获了包装后的慢病毒,病毒滴度为5×10^(7 )TU。感染HeLa细胞和BALB/c小鼠后,3D聚合酶表达量下降。本研究成功构建了慢病毒Lenti-sh3D,其能有效下调CVB 3D聚合酶的表达,为进一步抗CVB致病机制的研究奠定了基础。 | 闫碧莹 徐维祯 钟照华 | 2019 | 微生物与感染2019，14，2： | 0 |
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