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Soaking RNAi-mediated modification of Sf9 cells for baculovirus expression system by ectopic expression of Caenorhabditis elegans SID-1

  • Applied genetics and molecular biotechnology
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Abstract

RNA interference (RNAi) is a biological phenomenon that silences the expression of genes of interest. Passive double-stranded RNA (dsRNA) uptake has been uniquely observed in Caenorhabditis elegans due to the expression of systemic RNAi defective-1 (SID-1). We report that ectopic expression of CeSID-1 endows the Sf9 cells with a capacity for soaking RNAi. Soaking the Sf9-SID1 with dsRNA corresponding to either exogenous or endogenous target genes induced a significant decrease in the amount of mRNA or protein. These results enabled us to modify the target proteins of baculovirus expression vector system in both quantities and posttranslational modifications. The current low-cost and high-efficiency RNAi system is useful for high-throughput gene function analysis and mass production of recombinant protein.

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Acknowledgments

This work was supported in part by grants KAKENHI nos. 22248003, 22248004, and 23580077 from the Japan Society for the Promotion of Science. The cost of publication was supported in part by the Research Grant for Young Investigators of Faculty of Agriculture, Kyushu University.

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Authors and Affiliations

  1. Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, Fukuoka, 812-8581, Japan

    Jian Xu, Yudai Nagata, Hiroaki Mon, Zhiqing Li, Li Zhu, Takahiro Kusakabe & Jae Man Lee

  2. Laboratory of Insect Pathology and Microbial Control, Institute of Biological Control, Faculty of Agriculture, Graduate School, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka, 812-8581, Japan

    Kazuhiro Iiyama

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  1. Jian Xu
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  2. Yudai Nagata
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Correspondence to Jae Man Lee.

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Xu, J., Nagata, Y., Mon, H. et al. Soaking RNAi-mediated modification of Sf9 cells for baculovirus expression system by ectopic expression of Caenorhabditis elegans SID-1. Appl Microbiol Biotechnol 97, 5921–5931 (2013). https://doi.org/10.1007/s00253-013-4785-1

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