title: Electroporation of siRNA into mouse bone marrow-derived macrophages and dendritic cells.
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.
authors: Mikrobiologisches Institut - Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors: Isabel
authors: I
authors: Siegert
authors: Valentin
authors: V
authors: Schatz
authors: Alexander T
authors: AT
authors: Prechtel
authors: Alexander
authors: A
authors: Steinkasserer
authors: Christian
authors: C
authors: Bogdan
authors: Jonathan
authors: J
authors: Jantsch
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D003713:Dendritic Cells / Q000166:cytology / Q000378:metabolism*; D018274:Electroporation / Q000379:methods*; D055785:Gene Knockdown Techniques; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering