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[Specific suppression of GFP gene expression by short double stranded RNA in mammalian cells]
Yu-chen Huang1, Jiang Li, Chen Tan
1Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, P. R. China.
Background And 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.
Conclusion:
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.