Efficient delivery of siRNA targeted against human papillomavirus oncogenes.

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Finally, since RNAi directed against HPV18 E6/E7 has been reported to cause cellular senescence in HeLa cells (4), we monitored the typical senescence-associated spindly-cell and large-cell morphology of cells transfected with X-tremeGENE siRNA Transfection Reagent (Figure 1c). As expected, dramatic cell growth suppression was observed in HPV18 E6/E7 siRNA transfected HeLa cells together with the appearance of senescent cells. Similar effects were not observed in the mock-transfected HeLa cell control. Cells of the cell line 293--which does not harbor the HPV oncogenes--were used as a negative control for this experiment and were unaffected, suggesting that the RNAi approach was specific.
Conclusion
X-tremeGENE siRNA Transfection Reagent is an advanced product that allows efficient introduction of both DNA plasmid and duplex RNAs into cultured ceils at minimal toxicity. In the future, this reagent in conjunction with the HPV18 E6/E7 siRNA will be a valuable tool for the analysis of molecular pathways downstream from the HPV oncogenes.
Table I. HeLa cells were transfected using the X-tremeGENE siRNA
Transfection Reagent or Reagent A with either [beta]-gal expression
plasmid alone or together with siRNA. Cells were stained for
[beta]-gal activity at 24 h post-transfection and were microscopically
examined for signs of toxicity.

[beta]-Gal efficiency Toxicity
X-tremeGENE
[beta]-Gal 80-90% minor

Reagent A
[beta]-Gal none minor

X-tremeGENE
[beta]-Gal + siRNA 80-90% minor

Reagent A
[beta]-Gal + siRNA none minor
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