Institute of Pharmacy, Freie Universität Berlin, Berlin, Germany.
Methods Mol Biol. 2022;2521:157-171. doi: 10.1007/978-1-0716-2441-8_8.
Due to the lower risks of adverse effects, nonviral gene therapy is a suitable alternative to transfect cancer cells with a suicide gene to let them kill themselves by expressing toxic ribosome-inactivating proteins. Plasmids are stable and easy-to-produce vectors, but they have some disadvantages due to the bacterial backbone. Applying the minicircle technology, this problem can be solved with manageable effort in a well-equipped laboratory. With the described methodology, minicircle-DNA can be produced at low costs. The cell killing properties are monitored following transfection using the CytoSMART Omni system-a camera based live cell imaging device.
由于不良反应风险较低,非病毒基因治疗是一种合适的替代方法,可将自杀基因转染癌细胞,使其通过表达毒性核糖体失活蛋白而自我杀死。质粒是稳定且易于生产的载体,但由于其细菌骨架,它们存在一些缺点。通过应用微环技术,可以在设备齐全的实验室中以可控的努力解决这个问题。使用描述的方法,可以低成本地生产微环 DNA。使用 CytoSMART Omni 系统(一种基于摄像头的活细胞成像设备)在转染后监测细胞杀伤特性。