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Transfection of BmCPV genomic dsRNA in silkmoth-derived Bm5 cells: Stability and interactions with the core RNAi machinery

文献类型： 外文期刊

作者： Swevers, Luc 1 ; Kolliopoulou, Anna 1 ; Li, Zheng 2 ; Daskalaki, Maria 3 ; Verret, Frederic 3 ; Kalantidis, Kriton; 1 ;

作者机构： 1.Natl Ctr Sci Res Demokritos, Inst Biosci & Applicat, Athens 15342, Greece

2.South China Agr Univ, Coll Anim Sci, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Guangdong Engn Res Ctr Subtrop Sericulture & Mulb, Guangzhou 510642, Guangdong, Peoples R China

3.Univ Crete, Dept Biol, Iraklion 70013, Crete, Greece

4.Univ Ghent, Fac Biosci Engn, Dept Crop Protect, B-9000 Ghent, Belgium

关键词： Cytoplasmic polyhedrosis virus;BmCPV;dsRNA;RNAi;Bm5 cells

期刊名称：JOURNAL OF INSECT PHYSIOLOGY （ 影响因子：2.354； 五年影响因子：3.045 ）

ISSN： 0022-1910

年卷期： 2014 年 64 卷

页码：

收录情况： SCI

摘要： While several studies have been conducted to investigate the stability of dsRNA in the extracellular medium (hemolymph, gut content, saliva), little is known regarding the persistence of dsRNA once it has been introduced into the cell. Here, we investigate the stability of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) genomic dsRNA fragments after transfection into Bombyx-derived Bm5 cells. Using RT-PCR as a detection method, we found that dsRNA could persist for long periods (up to 8 days) in the intracellular environment. While the BmCPV genomic dsRNA was processed by the RNAi machinery, its presence had no effects on other RNAi processes, such as the silencing of a luciferase reporter by dsLuc. We also found that transfection of BmCPV genomic dsRNA could not establish a viral infection in the Bm5 cells, even when co-transfections were carried out with dsRNAs targeting Dicer and Argonaute genes, suggesting that the neutralization by RNAi does not play a role in the establishment of an in vitro culture system. The mechanism of the dsRNA stability in Bm5 cells is discussed, as well as the implications for the establishment for an in vitro culture system for BmCPV. (C) Elsevier Ltd. All rights reserved.

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