宿主基因TBL1XR1沉默对水疱性口炎病毒复制的影响

Effect of host TBL1XR1 silencing on replication of vesicular stomatitis virus

为揭示转导蛋白(β)样1X连接受体1(TBL1XR1)对水疱性口炎病毒(VSV)复制的影响,首先设计并构建沉默TBL1XR1的shRNA表达载体,并以其作为包装慢病毒的穿梭质粒,同骨架质粒共转染239T细胞后,成功包装出表达shTBL1XR1的慢病毒颗粒。以慢病毒感染小鼠巨噬细胞RAW264.7并建立TBL1XR1表达沉默的细胞系,同时构建不靶向任何基因的对照细胞系。最后,用VSV感染TBL1XR1沉默细胞系,并通过荧光定量PCR方法检测VSV的复制能力。结果发现,对TBL1XR1进行沉默能显著抑制VSV的复制,推测TBL1XR1及其相关复合物在VSV侵染过程中起到了一定的辅助作用。研究结果不仅为揭示VSV的致病机理奠定基础,也对VSV的疫病防控起到一定的指导作用。

Transducin(β)-like 1X-linked receptor 1(TBL1XR1)is a kind of important host factor,which regulates multiple processes in both physiological and pathological levels.This study aims to explore the influence of TBL1XR1on vesicular stomatitis virus(VSV)replication.We first designed and constructed shRNA expressing vector,which targets TBL1XR1gene to form a shuttle vector of lentivirus.The shuttle vector was then co-transfected lentivirus packaging vector into 293Tcell,and then recombinant lentivirus expressing shTBL1XR1was packaged.This was followed by virus infection to murine macrophage cell line RAW264.7.After puromycin selecting,RAW264.7cell line expressing shTBL1XR1was established,and at the same time RAW264.7control cell line was also established.Finally,the above established cell lines were tested by infecting VSV,and Q-PCR method was used to test the capacities of VSV replication.Based on the results,silencing TBL1XR1could significantly inhibit VSV replication,suggesting that TBL1XR1and its relevant complex are involved in the process of VSV infection.This study not only paves a way to uncover VSV infectious mechanisms,but also help to guide vesicular stomatitis(VS)prevention and control.