ShRNA expression plasmids generated by a novel method efficiently induce gene-specific knockdown in a silkworm cell line

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

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)

Abstract

RNAi knockdown by using shRNA expression plasmids is widely used to determine the function of individual genes in mammals. Here we developed a simple method to create an IR DNA in a U6 small nuclear RNA promoter-based parent vector using a single-stranded IR DNA with short hairpin structure and Bst DNA polymerase. Furthermore, we demonstrated that the shRNA expression plasmids constructed by our method effectively induced target-specific RNAi in the silkworm cell line. We also found that sequence preference in the silkworm cell line was much lower than in mammalian cells and shRNA-induced RNAi was influenced by the length of the stem region.

Original languageEnglish
Pages (from-to)173-179
Number of pages7
JournalMolecular Biotechnology
Volume41
Issue number2
DOIs
Publication statusPublished - 2009 Feb
Externally publishedYes

Keywords

  • Bombyx mori
  • Bst DNA polymerase
  • RNA interference
  • Short hairpin RNA
  • U6 promoter

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology

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