长链非编码RNA BTF3P11调控细胞代谢促进肠道病毒71型复制机制的研究

Enterovirus 71 (EV71), the major causative agent of hand, foot and mouth disease, can lead to severe cases. The differential expression of a series of long noncoding RNA (lncRNA) in host cells induced by EV71 infection can utilize lncRNA to hijack the metabolism of the host cells to promote the virus immune escape. We have found a key lncRNA-BTF3P11 associated with EV71 infection. To further clarify the molecular mechanism of lncRNA as intermediate molecular to promote EV71 replication, the models of BTF3P11 overexpression and knockout cell lines, and transgenic mice will be established using adeno-associated virus and CRISPR/Cas9 approaches to validate the influence of BTF3P11 on EV71 replication in vivo and in vitro. The transcriptome sequencing and ChIRP/MS will be carried out to search for the metabolic enzyme interacted with BTF3P11 and clarify the mechanism of the interaction of both. The eCLIP-Seq method will be used to confirm the combined domains and the key sequence of BTF3P11. This study will illustrate the new mechanism and function of BTF3P11 in regulating cell metabolism to promote EV71 replication.

肠道病毒71型（EV71）是引起儿童手足口病的主要病原体，可诱发重症。EV71感染宿主后可引起细胞内一系列长链非编码RNA（long noncoding RNA, lncRNA）的差异表达，病毒可借助lncRNA的调控功能劫持宿主细胞的新陈代谢，促进免疫逃避。为阐明lncRNA作为媒介分子促进EV71复制的分子机制，课题组通过RNA-Seq寻找到与EV71复制密切相关的lncRNA-BTF3P11为切入点，进一步拟采用腺相关病毒和CRISPR/Cas9技术建立BTF3P11过表达和敲除细胞系及转基因小鼠模型，体内外验证BTF3P11异常表达对EV71复制的影响；进而通过转录组测序结合ChIRP/MS和eCLIP-Seq技术寻找与BTF3P11相结合的代谢酶，并分析两者的互作机制，明确BTF3P11作用的结构域和序列。本研究旨在阐明BTF3P11通过调控细胞新陈代谢促进EV71复制的新机制。

肠道病毒71型（EV71）是引起儿童手足口病的主要病原体，可诱发重症。EV71感染宿主后可引起细胞内一系列长链非编码RNA（long noncoding RNA, lncRNA）的差异表达，病毒可借助lncRNA的调控功能劫持宿主细胞的新陈代谢，促进免疫逃避。为阐明lncRNA作为媒介分子促进EV71复制的分子机制，课题组通过RNA-Seq寻找到与EV71复制密切相关的lncRNA-BTF3P11为切入点，进一步采用腺相关病毒和CRISPR/Cas9技术建立BTF3P11过表达和敲除细胞系及转基因小鼠模型，体内外验证BTF3P11异常表达对EV71复制的影响；进而通过转录组测序寻找与BTF3P11相结合的氨基转移酶CrmG，CrmG参与细胞代谢影响EV71的复制，进一步通过BTF3P11突变体转染技术寻找到与CrmG作用的结构域和序列。本研究阐明了lncRNA BTF3P11通过调控细胞新陈代谢促进EV71复制的新机制。