1. Programming human dendritic cells with mRNA.
- Author
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Lee J, Boczkowski D, and Nair S
- Subjects
- Antigens biosynthesis, Antigens genetics, Antigens immunology, Cancer Vaccines genetics, Cancer Vaccines immunology, Cancer Vaccines metabolism, Cell Line, Cytokines biosynthesis, Cytokines genetics, Cytokines immunology, Humans, Neoplasms genetics, Neoplasms immunology, Neoplasms metabolism, Neoplasms therapy, RNA, Messenger genetics, RNA, Messenger immunology, RNA, Messenger metabolism, Toll-Like Receptors biosynthesis, Toll-Like Receptors genetics, Toll-Like Receptors immunology, Cell Engineering methods, Dendritic Cells cytology, Dendritic Cells immunology, Dendritic Cells metabolism, Gene Expression, RNA, Messenger chemistry, Transfection methods
- Abstract
Transfecting with in vitro transcribed, protein-encoding mRNA is a simple yet effective method to express high levels of the desired RNA-encoded proteins in primary cells. Cells can be transfected with antigen-encoding mRNA, which is translated into protein and is processed by the cellular antigen-processing pathway to generate antigen-presenting cells. Another elegant and increasingly popular application is to transfect cells with mRNA that encodes immune modulating molecules (cytokines, chemokines, toll-like receptors (TLRs), immune receptor ligands, immune receptor targeting antibodies) which, when translated into protein, can program cell behavior and/or function. In this chapter we describe an efficient method to deliver mRNA into human dendritic cells (DCs) by electroporation. This is currently the method of choice to deliver mRNA into antigen-presenting cells for generating vaccines for cancer immunotherapy.
- Published
- 2013
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