RNA干扰在甲壳动物抗病毒免疫中的作用研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2025-1438

摘要/Abstract

摘要：

RNA干扰（RNA interference, RNAi）是一种存在于真核生物中由小RNA介导的转录后基因沉默机制，也是甲壳类动物抗病毒免疫的核心通路和研究热点，本文系统梳理了甲壳类RNAi通路的基础研究进展、作为反向遗传学工具的应用现状及RNAi抗病毒疗法的发展态势。基础研究方面，对虾等甲壳类的Dicer-2切割病毒dsRNA产生siRNA，并与AGO2装配成RISC复合物特异性沉默病毒基因，但DNA病毒siRNA来源与VSR逃逸问题仍是研究的难点与重点。同时，对虾等也进化出环形RNA调控、JAK/STAT通路交叉对话及FoxO介导的非序列依赖性应答等辅助抗病毒机制。应用层面，RNAi已被广泛用于解析甲壳类在免疫、生殖、生长、应激、肢体再生及体色调控等生物学过程中基因的功能，同时也被用于抗病毒研究，通过注射、口服或浸泡递送靶向病毒必需基因的dsRNA，可显著降低对虾病毒感染的死亡率，但RNAi技术产业化推广仍面临VSR干扰、脱靶效应、dsRNA稳定性差、口服效率低及成本高等挑战。未来可聚焦以下研究方向：一是利用单细胞测序绘制RNAi时空图谱，解析siRNA的来源；二是开发pH响应纳米载体、益生菌活载体及病毒样颗粒等智能递送系统；三是构建“多靶点dsRNA鸡尾酒 + 抗VSR多肽 + 免疫增强剂”的联合疗法，协同抑制病毒复制并阻断VSR逃逸；四是探索CRISPR-Cas13、环形RNA及内源逆转录酶通路调控等新型RNA靶向技术，建立不依赖经典Dicer/AGO2的抗病毒策略。本文旨在为甲壳类病毒病的防控提供理论依据与实践参考。

关键词: 甲壳类动物, RNA干扰, 抗病毒免疫, 病毒RNAi抑制子, dsRNA递送, 水产养殖

Abstract:

RNA interference (RNAi) is a small RNA-mediated post-transcriptional gene silencing mechanism in eukaryotes and serves as a core antiviral immune pathway and research hotspot in crustaceans. This review systematically summarizes fundamental advances in the crustacean RNAi pathway, its current application as a reverse genetic tool, and the development trends of RNAi-based antiviral therapy. In terms of basic mechanisms, Dicer-2 in shrimp and other crustaceans cleaves viral dsRNA into siRNAs, which are then assembled with AGO2 into the RISC complex to specifically silence viral genes. However, the origins of siRNA derived from DNA viruses and the issue of VSR (viral suppressor of RNA silencing)-mediated escape remain challenging and critical research topics. Meanwhile, shrimp and other crustaceans have alsoevolved auxiliary antiviral mechanisms, including circular RNA regulation, crosstalk with the JAK/STAT pathway, and FoxO-mediated sequence-independent responses. On the application level, RNAi has been widely employed to elucidate gene functions involved in various biological processes,including immunity, reproduction, growth, stress response, limb regeneration, and pigmentation. It has also been used in antiviral research. Delivery of dsRNA targeting essential viral genes via injection, oral administration, or immersion significantly reduces viral infection-associated mortality in shrimp. Nevertheless, the industrial-scale promotion of RNAi technology still faces challenges, including VSR interference, off-target effects, poor dsRNA stability, low oral delivery efficiency, and high costs. Future efforts can focus on the following directions: using single-cell sequencing to map the spatiotemporal landscape of RNAi and elucidate the origin of siRNA; developing intelligent delivery systems such as pH-responsive nanocarriers, live probiotic vectors, and virus-like particles; constructing combination therapies “multi-target dsRNA cocktails, anti-VSR peptides, and immune enhancers” to synergistically inhibit viral replication and block VSR escape; and exploring novel RNA-targeting technologies, including CRISPR-Cas13, circular RNA, and endogenous reverse transcriptase pathway modulation, to establish antiviral strategies independent of the canonical Dicer/AGO2 machinery. This review aims to provide a theoretical basis and practical reference for the prevention and control of viral diseases in crustaceans.

Key words: crustaceans, RNA interference, antiviral immunity, viral suppressor of RNAi (VSR), dsRNA delivery, aquaculture

任晓桐, 卢永忠. RNA干扰在甲壳动物抗病毒免疫中的作用研究进展[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1438.

REN Xiao-tong, LU Yong-zhong. Research Progress in the Role of RNA Interference in the Antiviral Immunity of Crustaceans[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1438.

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