家蚕中影响RNA干扰效率的因子与dsRNA相互作用的研究

项目名称： 家蚕中影响RNA干扰效率的因子与dsRNA相互作用的研究

项目编号： No.31501898

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 动物学

项目作者： 刘吉升

作者单位： 广州大学

项目金额： 20万元

中文摘要： RNA干扰是一种转录后基因沉默的机制，它在双翅目的果蝇中的通路非常清楚，在鞘翅目的赤拟谷盗的效果非常明显；然而，在家蚕等鳞翅目昆虫中，RNA干扰的通路尚不明确，其成功率也非常低；这与其对dsRNA的吸收和传递、RNA干扰通路上核心基因的表达水平、昆虫体内的RNA降解酶、dsRNA引起免疫反应取代了RNA干扰反应等有关。近年来，我们对R2D2、dsRNA降解酶、Toll9等几个与dsRNA相互作用的因子进行了初步的研究；我们发现R2D2仅在少量的组织微量表达；dsRNA降解酶能在昆虫细胞表达并降解dsRNA；dsRNA能够抑制Toll9-1的转录水平，并上调持续表达Toll9-1的家蚕细胞中Dicer-2的转录水平；本项目将继续以R2D2、dsRNA降解酶、及Toll9为研究对象，以家蚕幼虫和家蚕Bm5细胞为材料，深入研究它们与dsRNA的相互作用、以及哪个因子可能影响家蚕的RNA干扰效率。

中文关键词： 家蚕；RNA干扰；双链RNA；因子；互相作用

英文摘要： RNAi is a mechanism of post-transcriptional regulation of gene expression, where the pathways are extensively studied in the dipteran model insect Drosophila melanogaster and the response is robust in the coleopteran model insect Tribolium castaneum. However, in Lepidoptera including the model insect Bombyx mori, the RNAi pathways are still not clear and a smaller rate of efficiency was reported. This refractoriness is due to the uptake and delivery of dsRNA, the expression levels of RNAi machinery components, presence of RNA degrading enzymes to clear dsRNA, and the immune response instead of RNAi response triggered by dsRNA. Recently, we have found that R2D2 was strictly and slightly expressed in silkmoth, dsRNase can digest dsRNA in cell culture, and dsRNA can inhibit the transcription of Toll9-1 and trigged the transcription of Dicer-2 in Toll9-1 persistent expression cell line. To gain insight into the factors that might determine the efficiency of RNAi, several factors, including R2D2, dsRNase and Toll9, will continue to be investigated. Their interactions with dsRNA and RNAi will be studied in the silkmoth B. mori and Bm5 cells.

英文关键词： Silkmoth;RNAi;dsRNA;factors;interactions