Weber Wilfried, Fussenegger Martin
Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.
Methods Mol Biol. 2011;692:235-49. doi: 10.1007/978-1-60761-971-0_17.
Synthetic quorum-sensing systems in mammalian cells has enabled the implementation of time- and distance-dependent bioprocesses, as well as the design of synthetic ecosystems emulating clinically important host-parasite interactions. In this chapter, we provide a detailed protocol of the design of a mammalian cell-to-cell signaling device and its integration into a mammalian quorum-sensing system for cell density-induced expression product genes. Cell-to-cell signaling is based on a sender cell, metabolically engineered for expression of alcohol dehydrogenase converting ethanol into acetaldehyde, and a receiver cell line for the dose-dependent translation of the acetaldehyde concentration into transgene expression by an acetaldehyde-responsive promoter. This protocol can be adapted easily to various cell types and transgenes for the design of versatile mammalian cell-based quorum-sensing systems.
哺乳动物细胞中的合成群体感应系统能够实现时间和距离依赖性生物过程，还能设计模拟临床上重要的宿主-寄生虫相互作用的合成生态系统。在本章中，我们提供了一个详细的方案，用于设计哺乳动物细胞间信号传导装置，并将其整合到哺乳动物群体感应系统中，以实现细胞密度诱导的表达产物基因。细胞间信号传导基于一个供体细胞，该供体细胞经过代谢工程改造以表达将乙醇转化为乙醛的乙醇脱氢酶，以及一个受体细胞系，该受体细胞系通过乙醛响应启动子将乙醛浓度进行剂量依赖性转化为转基因表达。该方案可轻松适用于各种细胞类型和转基因，以设计通用的基于哺乳动物细胞的群体感应系统。
