Enhancement of larval RNAi efficiency by over-expressing Argonaute2 in Bombyx mori.
Enhancement of larval RNAi efficiency by over-expressing Argonaute2 in Bombyx mori.
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DOI：
10.7150/ijbs.10235
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发表时间：
2015
影响因子：
9.2
通讯作者：
Huang Y
中科院分区：
文献类型：
--
作者：
Li Z;Zeng B;Ling L;Xu J;You L;Aslam AF;Tan A;Huang Y
RNA干扰已被描述为基因功能分析的强大遗传工具和害虫管理的一种有前途的方法。然而，由于不同的RNAi机制，不同昆虫的RNAi效率差异很大。鳞翅目昆虫包括大量的害虫和模式昆虫，如家蚕。然而，体内RNAi在鳞翅目中的成功仅有有限的报道，特别是在幼虫摄食最多、对寄主植物危害最大的阶段。提高鳞翅目幼虫RNAi的效率是发展基于RNAi的害虫管理策略的迫切需要。在本研究中，我们研究了保守的RNAi核心因子ArgAerte2(Ago2)在调节家蚕RNAi效率中的作用。我们证明了引入BmAgo2 dsRNA可以抑制BMN细胞和胚胎中的RNAi反应。此外，我们建立了几个转基因家蚕品系，以评估BmAgo2在幼虫RNAi中的作用。当使用dsRNA注射靶向DsRed时，过表达的BmAgo2显著促进了dsRNA介导的幼虫RNAi；当使用转基因shRNA表达靶向BmBlos2时，过表达的BmAgo2显著促进了shRNA介导的幼虫RNAi。我们的结果表明，BmAgo2参与了家蚕的RNAi，并为提高家蚕和鳞翅目昆虫的幼虫RNAi效率提供了一种有前途的途径。
RNA interference has been described as a powerful genetic tool for gene functional analysis and a promising approach for pest management. However, RNAi efficiency varies significantly among insect species due to distinct RNAi machineries. Lepidopteran insects include a large number of pests as well as model insects, such as the silkworm, Bombyx mori. However, only limited success of in vivo RNAi has been reported in lepidoptera, particularly during the larval stages when the worms feed the most and do the most harm to the host plant. Enhancing the efficiency of larval RNAi in lepidoptera is urgently needed to develop RNAi-based pest management strategies. In the present study, we investigate the function of the conserved RNAi core factor, Argonaute2 (Ago2), in mediating B. mori RNAi efficiency. We demonstrate that introducing BmAgo2 dsRNA inhibits the RNAi response in both BmN cells and embryos. Furthermore, we establish several transgenic silkworm lines to assess the roles of BmAgo2 in larval RNAi. Over-expressing BmAgo2 significantly facilitated both dsRNA-mediated larval RNAi when targeting DsRed using dsRNA injection and shRNA-mediated larval RNAi when targeting BmBlos2 using transgenic shRNA expression. Our results show that BmAgo2 is involved in RNAi in B. mori and provides a promising approach for improving larval RNAi efficiency in B. mori and in lepidopteran insects in general.
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