Li Zhiqian, Zeng Baosheng, Ling Lin, Xu Jun, You Lang, Aslam Abu F M, Tan Anjiang, Huang Yongping
1. Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. ; 2. University of Chinese Academy of Sciences, Beijing 100049, China.
1. Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Int J Biol Sci. 2015 Jan 5;11(2):176-85. doi: 10.7150/ijbs.10235. eCollection 2015.
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.
RNA干扰已被描述为一种用于基因功能分析的强大遗传工具，也是一种有前景的害虫治理方法。然而，由于不同的RNA干扰机制，RNA干扰效率在昆虫物种间差异显著。鳞翅目昆虫包括大量害虫以及模式昆虫，如家蚕（Bombyx mori）。然而，在鳞翅目中，仅报道了有限的体内RNA干扰成功案例，尤其是在幼虫阶段，此时幼虫进食最多且对寄主植物危害最大。迫切需要提高鳞翅目幼虫RNA干扰的效率，以制定基于RNA干扰的害虫治理策略。在本研究中，我们探究了保守的RNA干扰核心因子AGO2（Argonaute2）在介导家蚕RNA干扰效率中的作用。我们证明，导入家蚕AGO2双链RNA会抑制家蚕培养细胞（BmN细胞）和胚胎中的RNA干扰反应。此外，我们建立了多个转基因家蚕品系，以评估家蚕AGO2在幼虫RNA干扰中的作用。过表达家蚕AGO2显著促进了以DsRed为靶标时通过双链RNA注射介导的幼虫RNA干扰，以及以家蚕BmBlos2为靶标时通过转基因短发夹RNA表达介导的幼虫RNA干扰。我们的结果表明，家蚕AGO2参与家蚕的RNA干扰，并为提高家蚕及一般鳞翅目昆虫的幼虫RNA干扰效率提供了一种有前景的方法。
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