Linked Life Data

15449340

Source:http://linkedlifedata.com/resource/pubmed/id/15449340

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2004-9-30
pubmed:abstractText
Gene 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.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
1548-8969
pubmed:author
pubmed:copyrightInfo
Copyright 2004 Wiley-Liss, Inc.
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
301
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
820-7
pubmed:dateRevised
2009-10-5
pubmed:meshHeading
pubmed-meshheading:15449340-Animals, pubmed-meshheading:15449340-Cells, Cultured, pubmed-meshheading:15449340-Chick Embryo, pubmed-meshheading:15449340-DNA Primers, pubmed-meshheading:15449340-Electroporation, pubmed-meshheading:15449340-Evaluation Studies as Topic, pubmed-meshheading:15449340-Flow Cytometry, pubmed-meshheading:15449340-Gene Expression, pubmed-meshheading:15449340-Green Fluorescent Proteins, pubmed-meshheading:15449340-Luminescent Proteins, pubmed-meshheading:15449340-Microscopy, Fluorescence, pubmed-meshheading:15449340-Plasmids, pubmed-meshheading:15449340-RNA, Messenger, pubmed-meshheading:15449340-RNA, Small Interfering, pubmed-meshheading:15449340-RNA Interference, pubmed-meshheading:15449340-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:15449340-Transfection
pubmed:year
2004
pubmed:articleTitle
Application of RNA interference to chicken embryos using small interfering RNA.
pubmed:affiliation
Laboratory of Reproductive Physiology and Biotechnology, Department of Animal and Marine Bioresource Sciences, Kyushu University, Hakozaki, Fukuoka, Japan.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't