Evaluation of RNA interference in developing porcine granulosa cells using fluorescence reporter genes

Takuro Hirano, Nobuhiko Yamauchi, Fuminori Sato, Tomoki Soh, Masa Aki Hattori

研究成果: Contribution to journalArticle査読

10 被引用数 (Scopus)

抄録

Gene silencing using small interfering RNA (siRNA) may be useful for functional analyses of unidentified genes expressed during cell differentiation. The present study was performed to evaluate RNA interference (RNAi) in porcine granulosa cells stimulated with bovine FSH, by using two fluorescence reporter genes: a plasmid encoding green fluorescent protein (GFP) and a plasmid encoding red fluorescent protein (RFP). The siRNA targeting GFP mRNA sequence (GFP-siRNA) with both plasmids was introduced into cultured cells by lipofection. GFP- and RFP-expressing cells were observed under fluorescence microscopy and analyzed by flow cytometry. Strong fluorescence was observed after introduction of both plasmids into cells. The intensity of green fluorescence generated by GFP was greatly suppressed by introduction of GFP-siRNA, showing an approximate 70% decrease in the ratio of green to red fluorescence. Consequently, we concluded that gene silencing by siRNA can be used to analyze the functions of genes of interest during differentiation of porcine granulosa cells.

本文言語英語
ページ(範囲)599-603
ページ数5
ジャーナルJournal of Reproduction and Development
50
5
DOI
出版ステータス出版済み - 12 1 2004

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology

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