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 | |
| Publication status | Published - Feb 1 2013 |
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All Science Journal Classification (ASJC) codes
- Ecology, Evolution, Behavior and Systematics
- Agronomy and Crop Science
- Biochemistry, Genetics and Molecular Biology(all)
- Insect Science
Cite this
Coexpression of Escherichia coli RNase III in silkworm cells improves the efficiency of RNA interference induced by long hairpin dsRNAs. / Lee, Jae 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 journal › Article
}
TY - JOUR
T1 - Coexpression of Escherichia coli RNase III in silkworm cells improves the efficiency of RNA interference induced by long hairpin dsRNAs
AU - Lee, Jae Man
AU - Kojin, Yoshito
AU - Tatsuke, Tsuneyuki
AU - Mon, Hiroaki
AU - Miyagawa, Yoshitaka
AU - Kusakabe, Takahiro
PY - 2013/2/1
Y1 - 2013/2/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84872759713&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84872759713&partnerID=8YFLogxK
U2 - 10.1111/j.1744-7917.2012.01569.x
DO - 10.1111/j.1744-7917.2012.01569.x
M3 - Article
C2 - 23955827
AN - SCOPUS:84872759713
VL - 20
SP - 69
EP - 77
JO - Insect Science
JF - Insect Science
SN - 1672-9609
IS - 1
ER -