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pubmed-article:15449340pubmed:abstractTextGene silencing using small interfering RNA (siRNA) is not established in avian species. The present study was performed to evaluate RNA interference (RNAi) in the chicken embryo by using a dual fluorescence reporter assay, a plasmid encoding green fluorescent protein (GFP) and a plasmid encoding red fluorescent protein (RFP). The siRNA targeting the GFP mRNA sequence (GFP-siRNA) with both plasmids was introduced into cultured cells and whole embryos by lipofection and microelectroporation, respectively. GFP- and RFP-expressed cells and embryos were observed under fluorescent microscopy and analyzed by flow cytometer, and their mRNAs were analyzed by reverse transcription-PCR (RT-PCR). The strong fluorescence was observed by introducing both plasmids into cells. The intensity of the green fluorescence generated by GFP was greatly suppressed by introducing GFP-siRNA. RT-PCR analysis showed that introducing GFP-siRNA also decreased GFP mRNA levels. In contrast to GFP, the intensity of the red fluorescence generated by RFP and the RFP mRNA levels remained unchanged. In whole embryos, also, introducing GFP-siRNA specifically suppressed GFP expression, and the suppression was maintained for at least 72 h. Consequently, it was concluded that the gene silencing using siRNA is applicable to analyzing the function of genes of interest during avian embryogenesis.lld:pubmed
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pubmed-article:15449340pubmed:authorpubmed-author:ItoMakotoMlld:pubmed
pubmed-article:15449340pubmed:authorpubmed-author:KitagawaYasuo...lld:pubmed
pubmed-article:15449340pubmed:authorpubmed-author:SatoFuminoriFlld:pubmed
pubmed-article:15449340pubmed:authorpubmed-author:HattoriMasa-A...lld:pubmed
pubmed-article:15449340pubmed:authorpubmed-author:NakagawaTetsu...lld:pubmed
pubmed-article:15449340pubmed:copyrightInfoCopyright 2004 Wiley-Liss, Inc.lld:pubmed
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pubmed-article:15449340pubmed:pagination820-7lld:pubmed
pubmed-article:15449340pubmed:dateRevised2009-10-5lld:pubmed
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pubmed-article:15449340pubmed:year2004lld:pubmed
pubmed-article:15449340pubmed:articleTitleApplication of RNA interference to chicken embryos using small interfering RNA.lld:pubmed
pubmed-article:15449340pubmed:affiliationLaboratory of Reproductive Physiology and Biotechnology, Department of Animal and Marine Bioresource Sciences, Kyushu University, Hakozaki, Fukuoka, Japan.lld:pubmed
pubmed-article:15449340pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:15449340pubmed:publicationTypeComparative Studylld:pubmed
pubmed-article:15449340pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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