Pan Zhong-Hua, Gao Kun, Hou Cheng-Xiang, Wu Ping, Qin Guang-Xing, Geng Tao, Guo Xi-Jie
Sericultural Research Institute, Jiangsu University of Science and Technology, Zhenjiang 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, China; School of Biology & Basic Medical Science, Soochow University, Suzhou 215123, China.
Sericultural Research Institute, Jiangsu University of Science and Technology, Zhenjiang 212018, China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, China.
Gene. 2015 Jul 1;565(1):56-61. doi: 10.1016/j.gene.2015.03.069. Epub 2015 Apr 1.
Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is one of the major viral pathogens in silkworm. Its infection often results in significant losses to sericulture. Studies have demonstrated that RNAi is one of the important anti-viral mechanisms in organisms. In this study, three dsRNAs targeting the RNA-dependent RNA polymerase (RDRP) gene of BmCPV were designed and synthesized with 2'-F modification to explore their interference effects on BmCPV replication in silkworm larvae. The results showed that injecting dsRNA in the dosage of 4-6 ng per mg body weight into the 5th instar larvae can interfere with the BmCPV-RDRP expression by 93% after virus infection and by 99.9% before virus infection. In addition, the expression of two viral structural protein genes (genome RNA segments 1 and 5) was also decreased with the decrease of RDRP expression, suggesting that RNAi interference of BmCPV-RDRP expression could affect viral replication. The study provides an effective method for investigating virus replication as well as the virus-host interactions in the silkworm larvae using dsRNA.
家蚕细胞质型多角体病毒（BmCPV）是家蚕的主要病毒病原体之一。其感染常给养蚕业造成重大损失。研究表明，RNA干扰是生物体重要的抗病毒机制之一。本研究设计并合成了三种靶向BmCPV的RNA依赖RNA聚合酶（RDRP）基因的双链RNA（dsRNA），并进行了2'-F修饰，以探究它们对家蚕幼虫中BmCPV复制的干扰作用。结果显示，向5龄幼虫注射每毫克体重4 - 6 ng剂量的dsRNA，病毒感染后可使BmCPV-RDRP表达降低93%，病毒感染前可使其降低99.9%。此外，随着RDRP表达的降低，两个病毒结构蛋白基因（基因组RNA片段1和5）的表达也随之下降，这表明对BmCPV-RDRP表达的RNA干扰可影响病毒复制。该研究为利用dsRNA研究家蚕幼虫中的病毒复制以及病毒与宿主的相互作用提供了一种有效方法。
