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

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

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)599-603
Number of pages5
JournalJournal of Reproduction and Development
Volume50
Issue number5
DOIs
Publication statusPublished - Dec 1 2004

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granulosa cells
RNA interference
green fluorescent protein
reporter genes
small interfering RNA
fluorescence
swine
plasmids
gene silencing
fluorescence microscopy
cell differentiation
cultured cells
flow cytometry
genes
cells
cattle

All Science Journal Classification (ASJC) codes

  • Animal Science and Zoology

Cite this

Evaluation of RNA interference in developing porcine granulosa cells using fluorescence reporter genes. / Hirano, Takuro; Yamauchi, Nobuhiko; Sato, Fuminori; Soh, Tomoki; Hattori, Masa Aki.

In: Journal of Reproduction and Development, Vol. 50, No. 5, 01.12.2004, p. 599-603.

Research output: Contribution to journalArticle

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