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piggyBac transposon-derived targeting shRNA interference against the Bombyx mori nucleopolyhedrovirus (BmNPV)

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Abstract

The Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the most destructive diseases in silkworm, which has caused the main damage to sericulture industry. In this study, we developed a system of RNAi to prevent the BmNPV infection using the piggyBac transposon-derived targeting short hairpin RNA (shRNA) interference. The shRNAs targeting the genes of i.e.-1, lef-1, lef-2 and lef-3 of BmNPV were designed and used to inhibit the intracellular replication or multiplication of BmNPV in Bm cells. The highest activity was presented in the shRNA targeting the i.e.-1c of BmNPV, of which the inhibition rate reached 94.5 % in vitro. Further a stable Bm cell line of piggyBac transposon-derived targeting shRNA interference against BmNPV was established, which has a highly efficacious suppression on virus proliferation. These results indicated that the recombinant shRNA expression system was a useful tool for resistance to BmNPV in vitro. The approach by recombinant shRNAs opens a door of RNAi technology as a strategy that offering technically simpler, cheaper, and quicker gene knockdown for promising research and biotechnology application on silkworm lethal diseases.

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Acknowledgments

The work was supported by the National Basic Research Program of China under grand No. 2012CB114601 and the National Natural Science Foundation of China (No. NSFC:31372374/C1703), the Science and Technology Innovation Team of Zhejiang Province (No. 2010R50031) and Chinese Universities Scientific Fund.

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Authors and Affiliations

  1. Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, People’s Republic of China

    Fang Zhou, Rui-ting Chen, Yan Lu, Shuang Liang, Mei-xian Wang & Yun-gen Miao

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  1. Fang Zhou
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  2. Rui-ting Chen
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  3. Yan Lu
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  4. Shuang Liang
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  6. Yun-gen Miao
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Correspondence to Yun-gen Miao.

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Zhou, F., Chen, Rt., Lu, Y. et al. piggyBac transposon-derived targeting shRNA interference against the Bombyx mori nucleopolyhedrovirus (BmNPV). Mol Biol Rep 41, 8247–8254 (2014). https://doi.org/10.1007/s11033-014-3726-0

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