| Literature DB >> 20006615 |
Melanie Wiese1, Kirstin Castiglione, Michael Hensel, Ulrike Schleicher, Christian Bogdan, Jonathan Jantsch.
Abstract
Selective gene silencing by RNA interference (RNAi) is a valuable tool for the targeted manipulation of the development and/or function of cells. Using a fluorescein-labeled non-silencing siRNA duplex, we established a protocol for the electroporation of primary mouse macrophages which routinely yielded >95% transfected cells. Electroporation of siRNAs directed against MAPK1 and CD86 led to an efficient knock-down of cellular protein in bone marrow-derived mouse macrophages (BM-Mphi). Importantly, the electroporation procedure did not impair the viability of BM-Mphi, their ability to ingest or degrade E. coli or their capacity to express iNOS mRNA, to produce NO or to upregulate TNF and IL-6 mRNA in response to inflammatory stimuli such as LPS. Therefore, we propose that electroporation of silencing siRNAs into murine BM-Mphi is a highly efficient method to manipulate gene expression of BM-Mphi that does not cause toxicity or a non-specific alteration of macrophage biology. 2009 Elsevier B.V. All rights reserved.Entities:
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Year: 2009 PMID： 20006615 DOI： 10.1016/j.jim.2009.12.002
Source DB: PubMed Journal: J Immunol Methods ISSN： 0022-1759 Impact factor: 2.303