NSTI BioNano 2010

Transformation of gold nanoparticles into effective gene silencing vectors by oligoethylene glycol passivation

M.R. Papasani, D. Pokharel, A. Giri, V.V.R. Sai, P.J. Hrdlicka, R.A. Hill
University of Idaho, US

Keywords: gene knockdown, oligo-ethylene glycol, gold nanoparticales

Abstract:

Small interfering RNAs (siRNAs) are useful agents for gene knockdown. We have taken a systematic approach to investigate the use of GNP as delivery platforms for firefly luciferase (FFL) siRNAs and the effect of passivation and transfection agents on GNP-siRNA conjugate gene knockdown efficacy. 3T3-L1 cells expressing the FFL gene (3T3-L1-FFL cells) were transfected with GNP-FFL-siRNA, GNP-siRNA + oligoethylene glycol (GNP-siRNA-OEG) or GNP-siRNA + lipofectamine for 24 h and luciferase reporter assays were conducted. Three different control experiments were also conducted. 3T3-L1-FFL cells were incubated with (a) FFL-siRNA + lipofectamine (positive gene knockdown control) (b) scrambled siRNA + lipofectamine (negative gene knockdown control) and (c) GNP only. No gene knockdown was observed in cells incubated with GNP alone. The greatest efficacy was observed in the FFL-siRNAs + lipofectamine (positive gene knockdown control), FFL gene expression being reduced to 29% of the negative control level. GNP-siRNA-OEG induced high efficacy gene knockdown, 39% of negative control, while GNP-siRNA +lipofectamine showed intermediate gene knockdown efficacy, 65% of negative control. However, no knockdown was observed with GNP-siRNA. These results suggest that OEG passivation has a beneficial influence on cell uptake or intracellular routing of GNP-siRNA conjugates to the cytoplasm or both and thus improving siRNA efficacy.
 
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