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Electroporation of siRNA into Mouse Bone Marrow-Derived Macrophages and Dendritic Cells

Authors :
Valentin Schatz
Alexander T. Prechtel
Christian Bogdan
Alexander Steinkasserer
Isabel Siegert
Jonathan Jantsch
Source :
Methods in Molecular Biology ISBN: 9781461496311
Publication Year :
2014
Publisher :
Springer New York, 2014.

Abstract

Dendritic cells (DC) and macrophages (MΦ) play a pivotal role in antimicrobial defense, in the regulation of immune responses, and in maintaining tissue homeostasis. The analysis of DC and MΦ function relies on primary cells albeit these cells are known to be difficult to transfect. This makes the use of small interfering RNA (siRNA) for targeted manipulation of gene expression by RNA interference difficult. In the following chapter, we provide a detailed protocol for the successful transfer of siRNA via electroporation into a defined population of mouse bone marrow-derived MΦ or DC that does not cause toxicity to the myeloid cells or nonspecific alterations of their biological functions. Factors that influence the transfection and knockdown rate will be highlighted.

Details

ISBN :
978-1-4614-9631-1
ISBNs :
9781461496311
Database :
OpenAIRE
Journal :
Methods in Molecular Biology ISBN: 9781461496311
Accession number :
edsair.doi...........03f7be677fc1efb4847bb8254efc1f81
Full Text :
https://doi.org/10.1007/978-1-4614-9632-8_9