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Programming human dendritic cells with mRNA

Methods Mol Biol. 2013:969:111-25. doi: 10.1007/978-1-62703-260-5_8.

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.

Publication types

  • Review

MeSH terms

  • Antigens / biosynthesis
  • Antigens / genetics
  • Antigens / immunology
  • Cancer Vaccines / genetics
  • Cancer Vaccines / immunology
  • Cancer Vaccines / metabolism
  • Cell Engineering / methods*
  • Cell Line
  • Cytokines / biosynthesis
  • Cytokines / genetics
  • Cytokines / immunology
  • Dendritic Cells* / cytology
  • Dendritic Cells* / immunology
  • Dendritic Cells* / metabolism
  • Gene Expression*
  • Humans
  • Neoplasms / genetics
  • Neoplasms / immunology
  • Neoplasms / metabolism
  • Neoplasms / therapy
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / immunology
  • RNA, Messenger / metabolism
  • Toll-Like Receptors / biosynthesis
  • Toll-Like Receptors / genetics
  • Toll-Like Receptors / immunology
  • Transfection / methods*

Substances

  • Antigens
  • Cancer Vaccines
  • Cytokines
  • RNA, Messenger
  • Toll-Like Receptors