Abstract
RNA interference has been of great interest not only as a research tool to suppress gene expression but also as an emerging therapeutic strategy to silence disease genes. However, the therapeutic use of siRNA faces the in vivo delivery challenge. An alternative method that could potentially be used for siRNA delivery into primary immune cells for therapeutic purposes is an ex vivo route, whereby immune cells could be isolated from a patient, reprogrammed with siRNAs, and infused back into the same patient. This chapter describes siRNA delivery into human primary monocytes and dendritic cells using a standard electroporation technique. Dendritic cells occupy a central role in the immune system, orchestrating a wide repertoire of responses that span from the development of self-tolerance to the generation of protective CD8+ T cell immunity.
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References
Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811
Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotides RNAs mediate RNA interference in cultured mammalian cells. Nature 411:494–498
Sioud M (2004) Therapeutic siRNAs. Trends Pharmacol Sci 25:22–28
Behlke MA (2006) Progress towards in vivo use of siRNAs. Mol Ther 13:644–670
Sioud M (2005) On the delivery of small interfering RNAs into mammalian cells. Expert Opin Drug Deliv 2:639–651
Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392:245–252
Finn OJ (2008) Cancer immunology. N Engl J Med 358:2704–2715
Motz GT, Coukos G (2013) Deciphering and reversing tumor immune suppression. Immunity 39:61–73
Grütz G (2005) New insights into the molecular mechanism of interleukin-10-mediated immunosuppression. J Leukoc Biol 77:3–15
Munn DH, Mellor AL (2004) IDO expression by dendritic cells: tolerance and tryptophan catabolism. Nat Rev Immunol 4:762–774
Munn DH, Zhou M, Attwood JT, Bondarev I, Conway SJ, Marshall B, Brown C, Mellor AL (1998) Prevention of allogeneic fetal rejection by tryptophan catabolism. Science 281:1191–1193
Sioud M (2005) Induction of inflammatory cytokines and interferon responses by double-stranded and single-stranded siRNAs is sequence-dependent and requires endosomal localization. J Mol Biol 348:1079–1090
Flatekval GF, Sioud M (2009) Modulation of dendritic cell maturation and function with mono- and bifunctional small interfering RNAs targeting indoleamine 2,3-dioxygenase. Immunology, 128:e837–e848
Van Tendeloo VF, Ponsaerts P, Lardon F, Nijs G, Lenjou M, Van Broeckhoven C, Van Bockstaele DR, Berneman ZN (2001) Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. Blood 98:49–56
Acknowledgements
This work was supported by the Gene therapy program at the Norwegian Radium Hospital to M. Sioud.
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Sioud, M. (2015). Cytoplasmic Delivery of siRNAs to Monocytes and Dendritic Cells via Electroporation. In: Sioud, M. (eds) RNA Interference. Methods in Molecular Biology, vol 1218. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1538-5_6
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DOI: https://doi.org/10.1007/978-1-4939-1538-5_6
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Publisher Name: Humana Press, New York, NY
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