Ensenauer R, Hartl D, Vockley J, Roscher A A, Fuchs U
Children's Research Center, Dr. von Hauner Children's Hospital, Ludwig-Maximilians-Universität München, Munich, Germany.
Biotech Histochem. 2011 Aug;86(4):226-31. doi: 10.3109/10520291003675485. Epub 2010 Mar 18.
Magnetic force combined with magnetic nanoparticles recently has shown potential for enhancing nucleic acid delivery. Achieving effective siRNA delivery into primary cultured cells is challenging. We compared the utility of magnetofection with lipofection procedures for siRNA delivery to primary and immortalized mammalian fibroblasts. Transfection efficiency and cell viability were analyzed by flow cytometry and effects of gene knockdown were quantified by real-time PCR. Lipofectamine 2000 and magnetofection achieved high transfection efficiencies comparable to similar gene silencing effects of about 80%; the cytotoxic effect of magnetofection, however, was significantly less. Magnetofection is a reliable and gentle alternative method with low cytotoxicity for siRNA delivery into difficult to transfect cells such as mammalian fibroblasts. These features are especially advantageous for functional end point analyses of gene silencing, e.g., on the metabolite level.
磁力与磁性纳米颗粒相结合,最近已显示出增强核酸递送的潜力。将有效的小干扰RNA(siRNA)递送至原代培养细胞具有挑战性。我们比较了磁转染与脂质体转染方法在将siRNA递送至原代和永生化哺乳动物成纤维细胞中的效用。通过流式细胞术分析转染效率和细胞活力,并通过实时聚合酶链反应(PCR)对基因敲低的效果进行定量。Lipofectamine 2000和磁转染实现了高转染效率,与约80%的类似基因沉默效果相当;然而,磁转染的细胞毒性作用明显较小。磁转染是一种可靠且温和的替代方法,对于将siRNA递送至难以转染的细胞(如哺乳动物成纤维细胞)具有低细胞毒性。这些特性对于基因沉默的功能终点分析(例如在代谢物水平上)特别有利。