Gopalakrishnan Bagavathi, Wolff Jon
Mirus Bio Corporation, 505 S. Rosa Road, Madison, WI 53719, USA.
Methods Mol Biol. 2009;480:31-52. doi: 10.1007/978-1-59745-429-2_3.
Transfection is a powerful non-viral technology used to deliver foreign nucleic acids into eukaryotic cells, and is the method of choice for a variety of applications including studying the functional role of particular genes and the proteins they code for. By over-expressing genes to produce protein of interest and also by knocking down specific genes, researchers are able to accurately define the role of genes and the protein they encode in various cellular processes. Therefore, this powerful technology is a very vital component of the array of scientific research tools. However, the exact mechanism of action of transfection and also the numerous factors that influence the success of DNA or RNA delivery processes are not clearly understood. Hence, this chapter attempts to explain some of the popular cationic lipid/polymer-based transfection reagents for in vitro DNA/small inhibitory RNA (siRNA) delivery, mainly focusing on the protocols and critical factors to keep in mind to ensure successful delivery of nucleic acids into eukaryotic cells using these methods.
转染是一种强大的非病毒技术，用于将外源核酸导入真核细胞，是包括研究特定基因及其编码的蛋白质的功能作用在内的多种应用的首选方法。通过过表达基因以产生感兴趣的蛋白质，以及通过敲低特定基因，研究人员能够准确界定基因及其编码的蛋白质在各种细胞过程中的作用。因此，这项强大的技术是一系列科研工具中非常重要的组成部分。然而，转染的确切作用机制以及影响DNA或RNA递送过程成功的众多因素尚不清楚。因此，本章试图解释一些用于体外DNA/小干扰RNA（siRNA）递送的基于阳离子脂质/聚合物的常用转染试剂，主要关注使用这些方法将核酸成功递送至真核细胞时应牢记的方案和关键因素。
