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.
authors: Laboratory of Silkworm Sciences, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan.
authors: Jae Man
authors: JM
authors: Lee
authors: Yoshito
authors: Y
authors: Kojin
authors: Tsuneyuki
authors: T
authors: Tatsuke
authors: Hiroaki
authors: H
authors: Mon
authors: Yoshitaka
authors: Y
authors: Miyagawa
authors: Takahiro
authors: T
authors: Kusakabe
mesh_terms: D000818:Animals; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D055029:Inverted Repeat Sequences; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D043244:Ribonuclease III / Q000235:genetics* / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D012330:RNA, Double-Stranded; D043244:Ribonuclease III
keywords: Bombyx mori; RNA interference; RNase III; dsRNA; siRNA