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 from patients in 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 effects of siRNA electroporation on moDCs' viability, phenotype, migratory capacity, and ability to induce T-cell mediated 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 in electroporated moDCs. 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 an advantage especially in the light of clinical trials.
Department of Dermatology, University Hospital Erlangen, Hartmannstrasse 14, D-91052 Erlangen, Germany. alexander.prechtel@derma.imed.uni-erlangen.de
Alexander T
AT
Prechtel
Nadine M
NM
Turza
Alexandros A
AA
Theodoridis
Mirko
M
Kummer
Alexander
A
Steinkasserer
D002465:Cell Movement; D003713:Dendritic Cells / Q000276:immunology / Q000502:physiology*; D018274:Electroporation / Q000379:methods*; D020868:Gene Silencing; D006801:Humans; D016130:Immunophenotyping; D034904:Lamin Type A / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D012313:RNA / Q000737:chemistry / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D034904:Lamin Type A; D034741:RNA, Small Interfering; D012313:RNA