Zöllner Hannah, Hahn Stephan A, Maghnouj Abdelouahid
Labor für Molekulare Gastroenterologische Onkologie (MGO), Zentrum für Klinische Forschung (ZKF), Ruhr Universität Bochum, Bochum, Germany.
Methods Mol Biol. 2014;1095:177-90. doi: 10.1007/978-1-62703-703-7_15.
Deregulation of microRNAs (miRNAs) has been attributed to almost any human disease analyzed to date. This calls for models and experimental strategies for functional analyses of miRNAs enabling miRNA overexpression or suppression in target cell and/or tissues. Lentiviral vector (LV)-based technologies allow the long-term overexpression of miRNAs in nearly all cell types in an easy and rapid manner and are therefore popular tools for this application. In this chapter we describe the cloning of LV miRNA expression vectors as well as the production of virus particles for target cell infection and stable expression of miRNAs.
迄今为止，几乎所有已分析的人类疾病都与 microRNA（miRNA）失调有关。这就需要用于 miRNA 功能分析的模型和实验策略，以便在靶细胞和/或组织中实现 miRNA 的过表达或抑制。基于慢病毒载体（LV）的技术能够以简单快速的方式在几乎所有细胞类型中实现 miRNA 的长期过表达，因此是用于此应用的常用工具。在本章中，我们将描述 LV miRNA 表达载体的克隆，以及用于靶细胞感染和 miRNA 稳定表达的病毒颗粒的生产。
