白斑综合症病毒编码的lncRNA-8负调控对虾miR-34抗病毒免疫的机制研究

Long non-coding RNA (lncRNA) can be used as a competitive endogenous RNA (ceRNA) to regulate miRNA, but there are few studies on the interaction between lncRNA and miRNA in virus-host. The regulation of miRNA by lncRNA has not been involved especially in Marsupenaeus japonicus shrimp and white spot syndrome virus (WSSV). Previous studies have shown that shrimp miR-34 has antiviral immunity, and WSSV-encoded lncRNA-8 may be involved in the antiviral mechanism of miR-34 as a ceRNA. Therefore, based on the previous research, this project aims to clarify the effect of WSSV lncRNA-8 on viral infection, analyze the interaction between WSSV lncRNA-8 and miR-34, and further explore the mechanism of WSSV lncRNA-8 affecting viral infection through miR-34. Thus, the ceRNA regulatory network in host-virus interaction is established to reveal the mechanism of WSSV lncRNA-8 negatively regulating the anti-viral immunity of shrimp miR-34, providing new evidence for further understanding of the pathogenesis of WSSV and new ways for the prevention of shrimp white spot syndrome.

长链非编码RNA（lncRNA）可作为竞争性内源RNA（ceRNA）调控miRNA，但在病毒-宿主中lncRNA与miRNA的互作机制，研究较少。尤其在日本对虾与白斑综合症病毒（WSSV）中，lncRNA对miRNA的调控还未涉及。前期研究表明，对虾miR-34具有抗病毒免疫作用，WSSV编码的lncRNA-8可能作为ceRNA参与miR-34的抗病毒免疫机制。因此，本项目拟在前期研究基础上，通过分析WSSV lncRNA-8对病毒感染的影响、研究WSSV lncRNA-8与miR-34的互作关系并进一步探索WSSV lncRNA-8通过miR-34影响病毒感染的作用机理，来建立病毒宿主互作中的ceRNA调控网络，进而揭示WSSV lncRNA-8负调控miR-34抗病毒免疫的机制。为进一步认识病毒致病机制提供新证据，也为对虾白斑综合症的防控开辟新途径。

长链非编码RNA（lncRNA）可作为竞争性内源RNA（ceRNA）调控miRNA，但在病毒-宿主中lncRNA与miRNA的互作机制，研究较少。尤其在日本对虾与白斑综合症病毒（WSSV）中，lncRNA对miRNA的调控有待进一步研究。本项目利用RNAi、RNA 荧光原位杂交、靶基因体内外验证、流式细胞技术、miRNA过表达技术等分析了WSSV lncRNA-8对病毒感染的影响、研究了WSSV lncRNA-8与miR-34的互作关系并进一步探索了WSSV lncRNA-8通过miR-34影响病毒感染的作用机理。结果表明，WSSV lncRNA-8通过吸附具有抗病毒功能的对虾miR-34减弱其对靶基因EIF3A的调控，而EIF3A可以通过抑制细胞凋亡促进病毒复制，从而使WSSV lncRNA-8发挥促进病毒感染的功能。因此，本项目建立了对虾与WSSV互作过程中的lncRNA-miRNA-target mRNA调控网络，进而揭示了WSSV lncRNA-8负调控miR-34抗病毒免疫的机制。为进一步认识病毒致病机制提供新证据，也为对虾白斑综合症的防控开辟新途径。