Zhang Jun, He Qian, Zhang Chun-Dong, Chen Xiang-Yun, Chen Xue-Mei, Dong Zhan-Qi, Li Na, Kuang Xiu-Xiu, Cao Ming-Ya, Lu Cheng, Pan Min-Hui
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China; Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400016, China.
Antiviral Res. 2014 Apr;104:143-52. doi: 10.1016/j.antiviral.2014.01.017. Epub 2014 Jan 31.
Bombyx mori nucleopolyhedrovirus (BmNPV) is a major silkworm pathogen, causing substantial economic losses to the sericulture industry annually. We demonstrate a novel anti-BmNPV system expressing mature artificial microRNAs (amiRNAs) targeting the viral lef-11 gene. The mature amiRNAs inhibited the lef-11 gene in silkworm BmN-SWU1 cells. Antiviral assays demonstrated that mature amiRNAs silenced the gene and inhibited BmNPV proliferation efficiently. As constitutive overexpression of mature amiRNAs may induce acute cellular toxicity, we further developed a novel virus-induced amiRNA expression system. The amiRNA cassette is regulated by a baculovirus-induced fusion promoter. This baculovirus-induced RNA interference system is strictly regulated by virus infection, which functions in a negative feedback loop to activate the expression of mature amiRNAs against lef-11 and subsequently control inhibition of BmNPV replication. Our study advances the use of a regulatable amiRNA cassette as a safe and effective tool for research of basic insect biology and antiviral application.
家蚕核型多角体病毒（BmNPV）是一种主要的家蚕病原体，每年给养蚕业造成巨大经济损失。我们展示了一种新型的抗BmNPV系统，该系统表达靶向病毒lef-11基因的成熟人工微小RNA（amiRNA）。成熟的amiRNA在家蚕BmN-SWU1细胞中抑制lef-11基因。抗病毒试验表明，成熟的amiRNA使该基因沉默并有效抑制BmNPV增殖。由于成熟amiRNA的组成型过表达可能会诱导急性细胞毒性，我们进一步开发了一种新型的病毒诱导amiRNA表达系统。amiRNA盒由杆状病毒诱导的融合启动子调控。这种杆状病毒诱导的RNA干扰系统受到病毒感染的严格调控，其在负反馈回路中发挥作用，以激活针对lef-11的成熟amiRNA的表达，进而控制对BmNPV复制的抑制。我们的研究推动了可调控amiRNA盒作为一种安全有效的工具在基础昆虫生物学研究和抗病毒应用中的使用。
