Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.
Department of Microbiology, Faculty of Science, Chulalongkorn University, Thailand.
J Biotechnol. 2020 Sep 10;321:48-56. doi: 10.1016/j.jbiotec.2020.06.022. Epub 2020 Jun 29.
Outbreaks of diseases caused by yellow head virus (YHV) and white spot syndrome virus (WSSV) infection in shrimp have resulted in economic losses worldwide. DsRNA-mediated RNAi has been used to control these viruses, and the best target genes for efficient inhibition of YHV and WSSV are the protease and ribonuleotide reductase small subunit (rr2), respectively. However, one dsRNA can suppress only one virus, and therefore the production of multi-target dsRNA to effectively inhibit both YHV and WSSV is needed. In this study, plasmids pETpro-rr2_one stem and pETpro-rr2_two stems were constructed to produce two different forms of multi-target dsRNA in E. coli, which were designed specifically to both YHV protease and WSSV rr2 genes. The potency of each dsRNA in inhibiting YHV and WSSV and reducing shrimp death were investigated in L. vannamei. Shrimp were injected with the dsRNAs into the hemolymph before challenge with YHV or WSSV. The results showed that both dsRNAs could inhibit the viruses, however the one stem construct was more effective than the two stems construct when shrimp were infected with WSSV. This study establishes a potential strategy for dual inhibition of YHV and WSSV for further application in shrimp aquaculture.
虾类中由黄头病毒(YHV)和白斑综合征病毒(WSSV)感染引起的疾病爆发导致了全球范围内的经济损失。dsRNA 介导的 RNAi 已被用于控制这些病毒,而有效抑制 YHV 和 WSSV 的最佳靶基因分别是蛋白酶和核甙酸还原酶小亚基(rr2)。然而,一种 dsRNA 只能抑制一种病毒,因此需要生产多靶标 dsRNA 以有效抑制 YHV 和 WSSV。在这项研究中,构建了质粒 pETpro-rr2_one stem 和 pETpro-rr2_two stems,以在大肠杆菌中产生两种针对 YHV 蛋白酶和 WSSV rr2 基因的多靶标 dsRNA。在凡纳滨对虾中研究了每种 dsRNA 抑制 YHV 和 WSSV 以及减少虾死亡的能力。在 YHV 或 WSSV 攻毒前,将 dsRNA 注射到对虾的血淋巴中。结果表明,两种 dsRNA 均能抑制病毒,但当虾感染 WSSV 时,单茎结构比双茎结构更有效。这项研究为 YHV 和 WSSV 的双重抑制建立了一种潜在的策略,可进一步应用于虾类养殖。