Xu Jianyang, Han Fang, Zhang Xiaobo
School of Life Sciences, Xiamen University, Xiamen 361005, People's Republic of China; Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, People's Republic of China.
Antiviral Res. 2007 Feb;73(2):126-31. doi: 10.1016/j.antiviral.2006.08.007. Epub 2006 Sep 14.
White spot syndrome virus (WSSV) is a major shrimp pathogen causing large economic losses all over the world. So far, however, there is no efficient approach to control this virus. RNA interference (RNAi), which has been applied to silence virus genes in eukaryotic organisms. In this investigation, a specific 21bp short interfering RNA (vp28-siRNA) targeting a major envelope protein gene (vp28) of WSSV was used to induce gene silencing in vivo in Penaeus japonicus shrimp. It was found that the transcription and expression of vp28 gene were silenced by the sequence-specific vp28-siRNA. However, the RNAi effect disappeared or significantly weakened even if one-nucleotide mutation existed in the vp28-siRNA. As revealed by quantitative PCR, the vp28-siRNA caused a significant reduction in viral DNA production of WSSV-infected shrimp. When treated with the vp28-siRNA, WSSV-infected shrimp had a reduced mortality rate. After three injections of the vp28-siRNA, the virus was completely eradicated from WSSV-infected shrimp. These findings suggest that RNAi is capable of silencing sequence-specific genes of WSSV and might constitute a new therapeutic strategy for WSSV infection in shrimp.
白斑综合征病毒（WSSV）是一种主要的虾类病原体，在全球范围内造成巨大经济损失。然而，到目前为止，尚无有效的方法来控制这种病毒。RNA干扰（RNAi）已被应用于在真核生物中沉默病毒基因。在本研究中，一种靶向WSSV主要包膜蛋白基因（vp28）的特异性21bp短干扰RNA（vp28-siRNA）被用于在日本对虾体内诱导基因沉默。结果发现，序列特异性的vp28-siRNA使vp28基因的转录和表达沉默。然而，即使vp28-siRNA中存在一个核苷酸突变，RNAi效应也会消失或显著减弱。定量PCR结果显示，vp28-siRNA使WSSV感染虾的病毒DNA产量显著降低。用vp28-siRNA处理后，WSSV感染虾的死亡率降低。在三次注射vp28-siRNA后，病毒从WSSV感染的虾中被完全清除。这些发现表明，RNAi能够沉默WSSV的序列特异性基因，可能构成一种针对虾WSSV感染的新治疗策略。
