[Specific suppression of GFP gene expression by short double stranded RNA in mammalian cells].
OBJECTIVE
Double-stranded RNA (dsRNA) has been shown to interfere the function of specific genes in various species. The current study was designed to investigate whether short interfering RNAs(siRNAs) of green fluorescence protein(GFP) can inhibit the expression of GFP in mammalian cells.


METHODS
HeLa cells were grown in standard conditions and transfected with pEGFPC2 and long dsRNA or siRNAs. Fluoroscopy and fluoroscent spectrophotometer were used to evaluate impact of GFPds RNA, Cx26 siRNAs, and longer double-strand RNAs on GFP expression, and flow cytometry was used to evaluate the cell apoptosis.


RESULTS
Synthetic siRNAs can inhibite specific expression of GFP gene in human HeLa cell lines and siRNAs have no such specific effect. Longer double-stranded but no siRNAs can induce cell apoptosis.


CONCLUSIONS
These observations may open a path toward the use of siRNAs as a new tool for researching gene function in mammalian cells and a gene-specific therapeutic tool.
Huang
Yu-chen
YC
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, P. R. China.
Li
Jiang
J
Tan
Chen
C
Li
Xiao-ling
XL
Li
Gui-yuan
GY
D017209:Apoptosis; D000072259:Connexin 26; D017630:Connexins; D015870:Gene Expression / Q000187:drug effects*; D020868:Gene Silencing; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000494:pharmacology*
D017630:Connexins; C000607042:GJB2 protein, human; D008164:Luminescent Proteins; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D000072259:Connexin 26; D049452:Green Fluorescent Proteins