Small interfering RNA (siRNA) delivery into monocyte-derived dendritic cells by electroporation

Selective gene silencing by small interfering RNAs (siRNAs) has been shown to be an efficient method for the targeted manipulation of cellular functions. Chemical transfection reagents represent the current standard technique in siRNA duplex delivery into mammalian cells. However, when trying to manipulate cells isolated front patients fit clinical approaches, chemical agents might cause unwanted side effects. such as allergic reactions. or interfere with other cellular functions. In this study we describe electroporation as a suitable and efficient method for the delivery of siRNA into monocyte-derived dendritic cells (moDCs). Using a fluorescein-labeled non-silencing siRNA duplex as a model system, we carefully investigated the PC, migratory capacity, and ability to induce T-cell mediated effects of siRNA electroporation on moDCs' viability, phenotype immune responses. Finally, by using a standard duplex directed against the nuclear Lamins A and C we were able to demonstrate an efficient knockdown of a cellular messenger RNA fit electroporated moDiCs. We therefore propose siRNA electroporation into moDCs as an efficient method to manipulate DC function at large cell numbers without the use of chemical transfection reagents. This new approach represents,in advantage especially in the light of clinical trials. (c) 2006 Elsevier B.V All rights reserved.