Hüser Andreas, Hofmann Christian
DeveloGen AG, NL Berlin, Berlin, Germany.
Am J Pharmacogenomics. 2003;3(1):53-63.
Various methods have been developed to transfer and express genes in mammalian cells. Each method, whether virally, non-virally, or physically-based, has unique favorable features, but also drawbacks with respect to meeting desired and specific needs. Baculoviruses have been used since 1983 to express recombinant genes controlled by strong insect-virus promoters in their natural host (insect) cells. Today this is a well-established and easy to handle system for producing large quantities of recombinant proteins for numerous purposes. In 1995 it was first published that recombinant baculoviruses are able to deliver genes into mammalian cells. These genes are expressed provided that they are controlled by a promoter which is active in mammalian cells. Since then, various vector variants have been developed and numerous potential and meaningful applications have been described. It is not surprising that the use of baculovirus vectors as mammalian cell gene delivery vectors is constantly increasing and that the system is undergoing permanent improvements. Based on the convenience of the system to transfer genes into mammalian cells, baculoviruses can be applied in cell-based assays for drug screening to overcome the long periods of time required to generate stable cell lines. Baculovirus vectors are able to deliver very large DNA sequences into mammalian cells and vectors for toxic gene products can also be generated. In addition, baculoviruses are valuable tools for launching viral infection in cases where there is no appropriate cell culture system. Moreover, recent research has shown that the vectors can be applied in vivo. Depending on the design of the study, baculovirus vectors allow for sustained gene expression or are able to induce an immune response directed against the delivered and/or displayed gene product. The latter offers the opportunity to generate monoclonal antibodies against certain proteins that have failed by other means. In addition, it points to the potential usefulness of baculovirus vectors as new kinds of vaccines. Baculovirus vectors are therefore considered an enabling technology for various product opportunities.
人们已经开发出多种方法来在哺乳动物细胞中转移和表达基因。每种方法，无论是基于病毒、非病毒还是物理方式，都有其独特的优势，但在满足特定需求方面也存在缺点。自1983年以来，杆状病毒已被用于在其天然宿主（昆虫）细胞中表达由强昆虫病毒启动子控制的重组基因。如今，这是一个成熟且易于操作的系统，可用于大量生产多种用途的重组蛋白。1995年首次发表的研究表明，重组杆状病毒能够将基因导入哺乳动物细胞。只要这些基因由在哺乳动物细胞中具有活性的启动子控制，它们就能得到表达。从那时起，人们开发了各种载体变体，并描述了许多潜在且有意义的应用。毫不奇怪，杆状病毒载体作为哺乳动物细胞基因递送载体的使用量在不断增加，并且该系统也在不断改进。基于该系统将基因转移到哺乳动物细胞中的便利性，杆状病毒可应用于基于细胞的药物筛选试验，以克服生成稳定细胞系所需的长时间。杆状病毒载体能够将非常大的DNA序列导入哺乳动物细胞，还可生成用于有毒基因产物的载体。此外，在没有合适细胞培养系统的情况下，杆状病毒是引发病毒感染的宝贵工具。而且，最近的研究表明这些载体可在体内应用。根据研究设计，杆状病毒载体可实现持续的基因表达，或能够诱导针对所递送和/或展示的基因产物的免疫反应。后者为针对某些通过其他方法未能成功获得的蛋白质生成单克隆抗体提供了机会。此外，这也表明杆状病毒载体作为新型疫苗具有潜在的用途。因此，杆状病毒载体被认为是一种适用于多种产品机会的使能技术。
