Lagor William R, Johnston Julie C, Lock Martin, Vandenberghe Luk H, Rader Daniel J
Institute for Translational Medicine and Therapeutics, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
Methods Mol Biol. 2013;1027:273-307. doi: 10.1007/978-1-60327-369-5_13.
Adeno-associated viral vectors have proven to be excellent gene delivery vehicles for somatic overexpression. These viral vectors can efficiently and selectively target the liver, which plays a central role in lipoprotein metabolism. Both liver-expressed as well as non-hepatic secreted proteins can be easily examined in different mouse models using this approach. The dosability of adeno-associated viral (AAV) vectors, as well as their potential for long-term expression, makes them an excellent choice for assessing gene function in vivo. This section will cover the use of AAV to study lipoprotein metabolism-including vector design, virus production and purification, and viral delivery, as well as monitoring of transgene expression and resulting phenotypic changes. Practical information is provided to assist the investigator in designing, interpreting, and troubleshooting experiments.
腺相关病毒载体已被证明是用于体细胞过表达的优秀基因递送载体。这些病毒载体能够高效且选择性地靶向肝脏，而肝脏在脂蛋白代谢中起核心作用。使用这种方法，可以在不同的小鼠模型中轻松检测肝脏表达的蛋白以及非肝脏分泌的蛋白。腺相关病毒（AAV）载体的可定量性及其长期表达的潜力，使其成为在体内评估基因功能的理想选择。本节将介绍使用AAV研究脂蛋白代谢的相关内容，包括载体设计、病毒生产与纯化、病毒递送，以及转基因表达监测和由此产生的表型变化。还提供了实用信息，以协助研究人员设计、解释实验并解决实验故障。
