Mykhaylyk Olga, Sánchez-Antequera Yolanda, Vlaskou Dialekti, Hammerschmid Edelburga, Anton Martina, Zelphati Olivier, Plank Christian
Institute of Experimental Oncology and Therapy Research, Technische Universität München, München, Germany.
Methods Mol Biol. 2010;605:487-525. doi: 10.1007/978-1-60327-360-2_34.
In a magnetofection procedure, self-assembling complexes of enhancers like cationic lipids with plasmid DNA or small interfering RNA (siRNA) are associated with magnetic nanoparticles and are then concentrated at the surface of cultured cells by applying a permanent inhomogeneous magnetic field. This process results in a considerable improvement in transfection efficiency compared to transfection carried out with nonmagnetic gene vectors. This article describes how to synthesize magnetic nanoparticles suitable for nucleic acid delivery by liposomal magnetofection and how to test the plasmid DNA and siRNA association with the magnetic components of the transfection complex. Protocols are provided for preparing magnetic lipoplexes, performing magnetofection in adherent and suspension cells, estimating the association/internalization of vectors with cells, performing reporter gene analysis, and assessing cell viability. The methods described here can be used to screen magnetic nanoparticles and formulations for the delivery of nucleic acids by liposomal magnetofection in any cell type.
在磁转染过程中，阳离子脂质等增强剂与质粒DNA或小干扰RNA（siRNA）的自组装复合物与磁性纳米颗粒结合，然后通过施加永久不均匀磁场将其浓缩在培养细胞表面。与使用非磁性基因载体进行的转染相比，该过程导致转染效率有显著提高。本文介绍了如何通过脂质体磁转染合成适用于核酸递送的磁性纳米颗粒，以及如何测试质粒DNA和siRNA与转染复合物磁性成分的结合情况。提供了制备磁性脂质体、在贴壁细胞和悬浮细胞中进行磁转染、评估载体与细胞的结合/内化、进行报告基因分析以及评估细胞活力的方案。这里描述的方法可用于筛选磁性纳米颗粒和制剂，以通过脂质体磁转染在任何细胞类型中递送核酸。
