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

Research paper by Jian J Xu, Yudai Y Nagata, Hiroaki H Mon, Zhiqing Z Li, Li L Zhu, Kazuhiro K Iiyama, Takahiro T Kusakabe, Jae Man JM Lee

Indexed on: 08 Mar '13Published on: 08 Mar '13Published in: Applied Microbiology and Biotechnology



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.