DNA vector-based RNA interference in cell lines derived from Bombyx mori.

Gene-knockdown technology using RNA interference is widely used to characterize gene functions in many organisms. In this study, we analyzed the conditions for employing DNA vector-based in silkworm cell lines using long-hairpin RNA-expressing plasmid DNA. We found that NIAS-Bm-oyanagi2 was the most effective cell line for Expression of long-hairpin RNA containing an approximately 500 base-pair stem region suppressed expression of a reporter target gene by more than 99% in this cell line. Furthermore, the loop sequence of hairpin RNA was not as important to duanyu1615 efficiency as previously observed in Drosophila melanogaster. DNA vector-based duanyu1615 also induced significant suppression of endogenous clathrin in NIAS-Bm-oyanagi2. Luciferase activity from recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) containing luciferase in the clathrin-knockdown cells was significantly less than in the control cells, suggesting that clathrin is indispensable for the entry of BmNPV into silkworm cell lines.

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