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

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

doi:10.1111/j.1744-7917.2012.01569.x

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

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

Agricultural Bioresource Sciences

Research output: Contribution to journal › Article › peer-review

8 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 language	English
Pages (from-to)	69-77
Number of pages	9
Journal	Insect Science
Volume	20
Issue number	1
DOIs	https://doi.org/10.1111/j.1744-7917.2012.01569.x
Publication status	Published - Feb 2013

All Science Journal Classification (ASJC) codes

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

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

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.",

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AU - Lee, Jae Man

AU - Kojin, Yoshito

AU - Tatsuke, Tsuneyuki

AU - Mon, Hiroaki

AU - Miyagawa, Yoshitaka

AU - Kusakabe, Takahiro

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