Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.
Virus Res. 2013 Jan;171(1):252-6. doi: 10.1016/j.virusres.2012.11.012. Epub 2012 Nov 29.
RNAi activation in shrimp through dsRNA injection has been well demonstrated but oral delivery of dsRNA remains controversial. Therefore, this study was conducted to determine whether RNAi was induced in shrimp by ingestion of bacteria expressing dsRNA. We fed shrimp, Penaeus monodon and Litopenaeus vannamei, with inactivated bacteria expressing dsRNA specific to the shrimp genes (Rab7 and STAT). Forty-eight hours after 6 day-continuous feeding, the level of the targeted gene transcript was measured by semi-quantitative RT-PCR. Significant reduction of Rab7 as well as STAT transcript was observed when compared to that of control shrimp fed with bacteria containing the empty vector or bacteria expressing non-related dsRNA (GFP). Moreover, the suppression was detected not only in the hepatopancreas but also in the gills indicating the successful systemic induction of RNAi via oral delivery of dsRNA. Our results suggested that RNAi in shrimp could be triggered by ingestion of dsRNA expressing bacteria. Therefore, oral feeding is a practical approach which can be used to deliver dsRNA for further viral inhibition in farmed shrimp.
通过注射 dsRNA 激活虾体内的 RNAi 已得到充分证实,但 dsRNA 的口服递送仍存在争议。因此,本研究旨在确定通过摄入表达 dsRNA 的细菌是否能诱导虾体内产生 RNAi。我们用表达 Rab7 和 STAT 基因 dsRNA 的灭活细菌喂食对虾(斑节对虾和凡纳滨对虾)。连续喂食 6 天后 48 小时,通过半定量 RT-PCR 测量靶基因转录本的水平。与对照组相比,用含有空载体或表达非相关 dsRNA(GFP)的细菌喂养的虾体内 Rab7 和 STAT 转录本的水平明显降低。此外,这种抑制不仅在肝胰腺中检测到,在鳃中也检测到,表明通过口服递送 dsRNA 成功地在体内诱导了 RNAi。我们的结果表明,虾体内的 RNAi 可以通过摄入表达 dsRNA 的细菌来触发。因此,口服喂养是一种实用的方法,可以用来递送 dsRNA,以进一步抑制养殖虾中的病毒。