Summary
The ability to manipulate in vitro cultured dendritic cells (DCs) by transfection represents an attractive strategy to load these antigen-presenting cells with genetic material encoding various immunogenic epitopes. The gene transfer approach can also be applied to DCs with the aim of expressing immunologically active molecules such as cytokines, costimulatory molecules, or simply to transiently express proteins to perform cell biology studies. Available gene transfer technologies for DCs include both viral and non-viral vector-based approaches. In this chapter, we describe non-viral strategies of RNA transfection. Special emphasis is given to murine bone-marrow-derived DCs, since gene transfer to human DCs has been extensively described in the literature, especially in the context of cancer immunotherapy and other clinical applications. Methods to deliver small interfering RNA (siRNA) to DCs are described as well. Finally, the potential of exogenously delivered RNA to activate DCs is discussed and some practical advice to avoid DC activation is described.
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Acknowledgements
This work is supported by grants to PP from CNRS-INSERM, the Ministère de la Recherche et de la Technologie (ACI), La Ligue Nationale Contre le Cancer, and the Human Frontier of Science Program (HSFP). MC is supported by the Swiss National Science Foundation (SNF) and EKS is supported by the HSFP. The authors would like to thank Dr. Peter Ponsaerts (University of Antwerp, Belgium) for having provided the plasmid pSP64/GFP, Professor Eli Gilboa (Duke University, USA) for the plasmid pGEM4Z/GFP/A64, and Professor Matthias Hentze (EMBL, Heidelberg, Germany) for the plasmid pT3LUC(pA).
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Ceppi, M., Schmidt, E., Pierre, P. (2009). Genetic Modification of Murine Dendritic Cells by RNA Transfection. In: Reiner, N. (eds) Macrophages and Dendritic Cells. Methods in Molecular Biology™, vol 531. Humana Press. https://doi.org/10.1007/978-1-59745-396-7_10
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DOI: https://doi.org/10.1007/978-1-59745-396-7_10
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