Use of Bombyx mori U6 promoter for inducing gene-silencing in silkworm cells

Daisuke Ohtsuka, Tomonori Nakatsukasa, Ryosuke Fujita, Shin Ichiro Asano, Ken Sahara, Hisanori Bando

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Double-stranded (ds) RNA is a useful tool for sequence-specific gene silencing, now known as RNA interference (RNAi), in a variety of organisms. Studies on pol III promoter-based expression systems have been well documented showing them to be an effective tool for inducing RNAi in mammalian cells and Drosophila Schneider 2 (S2) cells, but not in silkworm cells. In this study, we developed a Bombyx mori U6 promoter (U6p)-based short hairpin RNA (shRNA) expression system for inducing RNAi in silkworm cells. First, a transient reporter assay showed that the transfection of plasmids containing U6p-driven shRNA-expression units (U6-shRNA units) caused a decrease in the expression of the target gene in BmN cells with the efficacy dependent on the length of the hairpin stem region. Subsequently, we demonstrated the ability of the U6-shRNA unit to confer resistance to Bombyx mori nucleopolyhedrovirus (BmNPV)-resistance on BmN cells; i.e., the viral titer in the culture medium of BmN cells transfected with U6-shRNA (with the sequence of an essential viral gene)-expression plasmids was decreased to 30% of the control at 48 hr post-infection. These results suggested that the U6-shRNA expression system was a useful tool for RNAi in silkworm cells.

Original languageEnglish
Pages (from-to)125-131
Number of pages7
JournalJournal of Insect Biotechnology and Sericology
Volume77
Issue number3
Publication statusPublished - Oct 1 2008
Externally publishedYes

Fingerprint

Bombyx
Gene Silencing
gene silencing
Bombyx mori
silkworms
RNA
Genes
promoter regions
small interfering RNA
Small Interfering RNA
RNA Interference
RNA interference
cells
plasmids
Plasmids
Bombyx mori nucleopolyhedrovirus
Nucleopolyhedrovirus
Gene Expression
Double-Stranded RNA
Viral Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Business, Management and Accounting(all)
  • Agricultural and Biological Sciences(all)
  • Insect Science
  • Industrial and Manufacturing Engineering

Cite this

Ohtsuka, D., Nakatsukasa, T., Fujita, R., Asano, S. I., Sahara, K., & Bando, H. (2008). Use of Bombyx mori U6 promoter for inducing gene-silencing in silkworm cells. Journal of Insect Biotechnology and Sericology, 77(3), 125-131.

Use of Bombyx mori U6 promoter for inducing gene-silencing in silkworm cells. / Ohtsuka, Daisuke; Nakatsukasa, Tomonori; Fujita, Ryosuke; Asano, Shin Ichiro; Sahara, Ken; Bando, Hisanori.

In: Journal of Insect Biotechnology and Sericology, Vol. 77, No. 3, 01.10.2008, p. 125-131.

Research output: Contribution to journalArticle

Ohtsuka, D, Nakatsukasa, T, Fujita, R, Asano, SI, Sahara, K & Bando, H 2008, 'Use of Bombyx mori U6 promoter for inducing gene-silencing in silkworm cells', Journal of Insect Biotechnology and Sericology, vol. 77, no. 3, pp. 125-131.
Ohtsuka, Daisuke ; Nakatsukasa, Tomonori ; Fujita, Ryosuke ; Asano, Shin Ichiro ; Sahara, Ken ; Bando, Hisanori. / Use of Bombyx mori U6 promoter for inducing gene-silencing in silkworm cells. In: Journal of Insect Biotechnology and Sericology. 2008 ; Vol. 77, No. 3. pp. 125-131.
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