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

Kosuke Fujita, Aki Sagisaka, Kazuya Tomimoto, Jun Ishibashi, Shigeo Imanishi, Minoru Yamakawa, Hiromitsu Tanaka

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

Gene-knockdown technology using RNA interference (RNAi) is widely used to characterize gene functions in many organisms. In this study, we analyzed the conditions for employing DNA vector-based RNAi in silkworm cell lines using long-hairpin RNA-expressing plasmid DNA. We found that NIAS-Bm-oyanagi2 was the most effective cell line for RNAi. 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 RNAi efficiency as previously observed in Drosophila melanogaster. DNA vector-based RNAi 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.

Original languageEnglish
Pages (from-to)2026-2031
Number of pages6
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number9
DOIs
Publication statusPublished - 2009

Keywords

  • Bmnpv
  • Bombyx mon
  • Cell lines
  • Clathrin
  • RNA interference

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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