Keller Andrea-Anneliese, Maeß Marten B, Schnoor Michael, Scheiding Berith, Lorkowski Stefan
Institute of Nutrition, Friedrich Schiller University Jena, Jena, Germany.
Competence Cluster for Nutrition and Cardiovascular Health (nutriCARD) Halle-Jena-Leipzig, Leipzig, Germany.
Methods Mol Biol. 2018;1784:187-195. doi: 10.1007/978-1-4939-7837-3_18.
Transfection is defined as the transfer of foreign nucleic acids into cells. In general, transfection may achieve either overexpression of a gene by the transfer of plasmid DNA or suppression of gene expression by RNA interference after transfer of small interfering RNA. Both approaches allow for the detailed investigation of the function of a particular gene product or mechanisms of gene regulation. Macrophages are considered as hard-to-transfect cells, as they have evolved to recognize foreign nucleic acids and to initiate an immune response to these molecules. The presented electroporation protocol provides an effective tool to efficiently transfect human THP-1 macrophages with siRNA or plasmid DNA while avoiding macrophage activation.
转染被定义为将外源核酸导入细胞。一般来说,转染可通过质粒DNA的导入实现基因的过表达,或在导入小干扰RNA后通过RNA干扰实现基因表达的抑制。这两种方法都有助于对特定基因产物的功能或基因调控机制进行详细研究。巨噬细胞被认为是难以转染的细胞,因为它们已经进化到能够识别外源核酸并对这些分子启动免疫反应。本文介绍的电穿孔方案提供了一种有效的工具,可在避免巨噬细胞激活的同时,用小干扰RNA或质粒DNA高效转染人THP-1巨噬细胞。