RRC ID 29699
Author Lee JM, Kojin Y, Tatsuke T, Mon H, Miyagawa Y, Kusakabe T.
Title Coexpression of Escherichia coli RNase III in silkworm cells improves the efficiency of RNA interference induced by long hairpin dsRNAs.
Journal Insect Sci.
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.
Volume 20(1)
Pages 69-77
Published 2013-2
DOI 10.1111/j.1744-7917.2012.01569.x
PMID 23955827
MeSH Animals Bombyx / genetics* Bombyx / metabolism Escherichia coli / enzymology* Escherichia coli / genetics Escherichia coli Proteins / genetics* Escherichia coli Proteins / metabolism Gene Expression Inverted Repeat Sequences RNA Interference* RNA, Double-Stranded / chemistry RNA, Double-Stranded / genetics* RNA, Double-Stranded / metabolism Ribonuclease III / genetics* Ribonuclease III / metabolism
IF 2.71
Times Cited 8