Molecular Discovery Research, GlaxoSmithKline R&D, UE0433, 709 Swedeland Road, King of Prussia, PA 19406, USA +1 610 270 7602 ; +1 610 270 7359 ;
Expert Opin Drug Discov. 2009 Mar;4(3):243-56. doi: 10.1517/17460440902751599.
Adenovirus, retrovirus and lentivirus-based vectors, originally engineered and optimized for in vivo and ex vivo gene therapy, have become increasingly useful for viral-mediated gene delivery to support in vitro cell-based assays. Viral vectors underpin functional genomics screening of cDNA, shRNA and aptamer libraries, are used for a variety of target validation studies and importantly, for high-throughput cell-based drug discovery and compound profiling assays. The baculovirus/insect cell expression system had gained prevalence as a tool for recombinant protein production when it was observed that recombinant baculovirus vectors too could serve as efficient gene delivery vehicles for a wide range of mammalian cells. Although the use of baculovirus vectors in vivo has lagged behind retroviral, adenoviral and lentiviral vectors, they have gained prominence for development of in vitro cell-based assays due to the ease of generation, broad host range and excellent biosafety profile. There is an increasing emphasis on cell-based assays in high-throughput automated drug discovery laboratories and a variety of commercially available viral-vectors can be used for supporting these assays.
We compare and contrast the current viral-mediated gene delivery vector systems and highlight their suitability for cell-based drug discovery assays.
Viral-mediated gene delivery is increasingly being used in support of genome scale target validation studies and cell-based assay development for specific drug target genes such as ion channels, G protein-coupled receptors and intracellular enzymes. The choice of a delivery system over another for a particular application is largely dictated by the cell types and cell lines in use, virus cellular tropism, assay throughput, safety requirements and ease/cost of reagent generation.
腺病毒、逆转录病毒和慢病毒载体最初是为体内和体外基因治疗而设计和优化的，现在越来越多地用于病毒介导的基因传递，以支持体外细胞为基础的测定。病毒载体为 cDNA、shRNA 和适体文库的功能基因组筛选提供了支持，用于各种靶标验证研究，而且重要的是，还用于高通量基于细胞的药物发现和化合物分析测定。当观察到重组杆状病毒载体也可以作为广泛的哺乳动物细胞的有效基因传递载体时，杆状病毒/昆虫细胞表达系统已成为重组蛋白生产的工具而得到普及。尽管杆状病毒载体在体内的应用落后于逆转录病毒、腺病毒和慢病毒载体，但由于其易于生成、宿主范围广泛和出色的生物安全性，它们在体外细胞为基础的测定中得到了重视。在高通量自动化药物发现实验室中，基于细胞的测定的重要性日益增加，并且有多种市售的病毒载体可用于支持这些测定。
我们比较和对比当前的病毒介导的基因传递载体系统，并强调它们对基于细胞的药物发现测定的适用性。
病毒介导的基因传递越来越多地用于支持基因组规模的靶标验证研究和特定药物靶标基因（如离子通道、G 蛋白偶联受体和细胞内酶）的基于细胞的测定的开发。对于特定应用，选择一种传递系统而不是另一种传递系统主要取决于所使用的细胞类型和细胞系、病毒的细胞嗜性、测定的通量、安全性要求以及试剂生成的难易程度和成本。
