Coexpression of Escherichia coli RNase III in silkworm cells improves the efficiency of RNA interference induced by long hairpin dsRNAs

Man Lee, Yoshito Kojin, Tsuneyuki Tatsuke, Hiroaki Mon, Yoshitaka Miyagawa, Takahiro Kusakabe

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Long hairpin dsRNA transcribed from chromosomal DNA can induce RNA interference in Bombyx mori cells, although its gene silencing efficiency is lower than that of exogenously introduced double-stranded RNAs (dsRNAs). To solve this problem, we monitored the nuclear cytoplasmic translocation of the transcribed hairpin dsRNA and analyzed the processing efficiency into mature small interfering RNA (siRNA). Northern blot analysis revealed that the transcribed hairpin dsRNAs were spliced and transported into the cytoplasm, but were not effectively diced into siRNAs. Interestingly, RNAi with hairpin dsRNAs from genome-integrated IR transgene was stimulated by the coexpression of Escherichia coli RNase III, although this exogenous enzyme seemed to bring about nonspecific cleavage of cellular mRNA.

Original languageEnglish
Pages (from-to)69-77
Number of pages9
JournalInsect Science
Volume20
Issue number1
DOIs
Publication statusPublished - Feb 1 2013

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Ribonuclease III
Bombyx
Double-Stranded RNA
double-stranded RNA
RNA Interference
silkworms
RNA interference
Escherichia coli
RNA
Genes
Gene Silencing
cells
Transgenes
Northern Blotting
Small Interfering RNA
Cytoplasm
Genome
gene silencing
cytoplasm
Bombyx mori

All Science Journal Classification (ASJC) codes

  • Insect Science
  • Ecology, Evolution, Behavior and Systematics
  • Agronomy and Crop Science
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Coexpression of Escherichia coli RNase III in silkworm cells improves the efficiency of RNA interference induced by long hairpin dsRNAs. / Lee, Man; Kojin, Yoshito; Tatsuke, Tsuneyuki; Mon, Hiroaki; Miyagawa, Yoshitaka; Kusakabe, Takahiro.

In: Insect Science, Vol. 20, No. 1, 01.02.2013, p. 69-77.

Research output: Contribution to journalArticle

Lee, Man ; Kojin, Yoshito ; Tatsuke, Tsuneyuki ; Mon, Hiroaki ; Miyagawa, Yoshitaka ; Kusakabe, Takahiro. / Coexpression of Escherichia coli RNase III in silkworm cells improves the efficiency of RNA interference induced by long hairpin dsRNAs. In: Insect Science. 2013 ; Vol. 20, No. 1. pp. 69-77.
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