Saxena Vaibhav
Sancilio & Company, Inc., 2129 N. Congress Avenue, Riviera Beach, FL, 33404, USA.
Methods Mol Biol. 2017;1522:245-250. doi: 10.1007/978-1-4939-6591-5_19.
DNA-binding cationic liposomes are routinely employed as one of the major non-viral transfecting agents delivering DNA and other genes inside the cells. Cationic liposomes when complexed with DNA form a strong positively charged liposome-DNA complex or lipoplex. The chapter discusses primarily the major preparation technique for cationic liposomes and its physical characterization, with a focus on SYBR Green-I dye exclusion assay and DNA encapsulation enhancement by freeze-thaw technique. SYBR Green-I dye exclusion assay is a technique to determine the total amount of liposomal lipids required to bind a unit weight of DNA, which is critical for transfection experiments. Freeze-thaw technique on the other hand is one of the major techniques to improve DNA encapsulation in cationic liposomes.
DNA结合阳离子脂质体通常被用作主要的非病毒转染剂之一，用于将DNA和其他基因递送至细胞内部。阳离子脂质体与DNA复合时会形成带强正电荷的脂质体-DNA复合物或脂质纳米粒。本章主要讨论阳离子脂质体的主要制备技术及其物理特性，重点是SYBR Green-I染料排除测定法以及通过冻融技术增强DNA包封。SYBR Green-I染料排除测定法是一种确定结合单位重量DNA所需脂质体脂质总量的技术，这对转染实验至关重要。另一方面，冻融技术是提高阳离子脂质体中DNA包封率的主要技术之一。
