Stearylated octaarginine and artificial virus-like particles for transfection of siRNA into primary rat neurons.
RNA interference (RNAi) provides a powerful experimental tool for sequence-specific gene silencing, allowing efficient analysis of gene function in a multitude of cell types. However, application of RNAi in primary mammalian neurons has been limited by low-transfection efficiency and considerable toxicity of conventional transfection methods. In this study, we evaluated a peptide-mediated and a polymer/lipid-based cellular delivery method for siRNA into rat primary neurons and compared the results with a commonly used liposomal transfection reagent. Stearylated octaarginine (Stearyl-R8) was used as polypeptide and artificial virus-like particles (AVPs) were used as a combined liposomal-polymeric vector, since both reagents have been previously shown to successfully transfect DNA into cell lines. Stearyl-R8 and AVPs both promoted siRNA transfection into primary hippocampal neurons via the endosomal pathway. SiRNA-mediated gene silencing could be effectively induced in primary neuron cultures. In comparison with the commonly used cationic liposome transfection agent, both novel reagents were less detrimental to cell metabolic activity. We conclude that these novel transfection methods yield performances comparable to cationic liposome-mediated transfection for siRNA, while being less cytotoxic in primary neurons. Stearyl-R8 and AVPs may therefore represent novel and more cost-efficient alternatives to conventional siRNA-transfection reagents.
Tönges
Lars
L
Department of Neurology, Georg-August-University Göttingen, Faculty of Medicine, Waldweg, Göttingen, Germany
Lingor
Paul
P
Egle
Roman
R
Dietz
Gunnar P H
GP
Fahr
Alfred
A
Bähr
Mathias
M
D000818:Animals; D001120:Arginine; D001483:Base Sequence; D002470:Cell Survival; D002478:Cells, Cultured; D006624:Hippocampus / Q000502:physiology*; D008055:Lipids; D009474:Neurons / Q000502:physiology*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D009843:Oligoribonucleotides; D010902:Pituitary Gland / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D013229:Stearic Acids; D014162:Transfection
D008055:Lipids; C086724:Lipofectamine; D009842:Oligopeptides; D009843:Oligoribonucleotides; D034741:RNA, Small Interfering; D013229:Stearic Acids; C031183:stearic acid; D001120:Arginine