Inhibition of viral gene expression and replication in mosquito cells by dsRNA-triggered RNA interference
Inhibition of viral gene expression and replication in mosquito cells by dsRNA-triggered RNA interference
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DOI：
10.1006/mthe.2002.0652
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发表时间：
2002-08-01
影响因子：
12.4
通讯作者：
Morgan, RA
中科院分区：
文献类型：
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作者：
Caplen, NJ;Zheng, ZL;Morgan, RA
蚊子将许多病毒病原体传播给人类，包括每年影响约5000万人的登革热病毒。抑制昆虫宿主内的病毒基因表达可用于阻断病毒复制和随后将病原体传播给人类。由双链RNA（dsRNA）触发的自然发生的基因沉默机制，RNA干扰（RNAi），最近在包括果蝇在内的许多物种中被描述。为了确定dsRNA触发的RNAi是否存在于蚊子细胞中，我们使用白纹伊蚊C6/36细胞，并研究阻断病毒基因表达和复制的可行性，我们使用两种蚊媒病毒，Semliki森林病毒（SFV）和血清型1登革热病毒（DEN 1）。我们证明了dsRNA可以特异性地抑制C6/36细胞中来自质粒和SFV复制子的转基因表达，并且可以显著地改变DEN 1 RNA和病毒复制的动力学。由dsRNA介导的抑制是序列特异性的，并且等于或上级于由反义单链RNA（ssRNA）诱导的抑制。这项研究证明了dsRNA触发的蚊子细胞中基因表达和病毒复制的抑制，并表明这种机制可用于阻断昆虫宿主内的病原体复制，从而阻断疾病传播。
Mosquitoes transmit numerous viral pathogens to humans including dengue virus which affects similar to 50 million individuals per year. Inhibition of viral gene expression within an insect host could be used to block virus replication and subsequent transmission of the pathogen to humans. A naturally occurring gene silencing mechanism triggered by double-stranded RNA (dsRNA), RNA interference (RNAi), has recently been described in a number of species including Drosophila. To ascertain if dsRNA-triggered RNAi is present in mosquito cells, we used Aedes albopictus C6/36 cells, and to investigate the feasibility of blocking viral gene expression and replication, we used two mosquito-borne viruses, Semliki Forest virus (SFV) and the serotype 1 dengue virus (DEN1). We demonstrate that dsRNA can specifically inhibit transgene expression in C6/36 cells from both plasmid and SFV replicons and can significantly modify the kinetics of DEN1 RNA and virus replication. The inhibition mediated by dsRNA was sequence-specific and either equal or superior to that induced by antisense single-stranded RNA (ssRNA). This study demonstrates dsRNA-triggered inhibition of gene expression and virus replication in mosquito cells and suggests that this mechanism could be used to block pathogen replication within an insect host and, thus, block disease transmission.