Saxena Pratik, Bojar Daniel, Fussenegger Martin
Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, Basel, CH-4058, Switzerland.
Faculty of Science, University of Basel, Mattenstrasse 26, Basel, CH-4058, Switzerland.
Methods Mol Biol. 2017;1651:263-273. doi: 10.1007/978-1-4939-7223-4_19.
Synthetic biology, the synthesis of engineering and biology, has rapidly matured and has dramatically increased the complexity of artificial gene circuits in recent years. The deployment of intricate synthetic gene circuits in mammalian cells requires the establishment of very precise and orthogonal control of transgene expression. In this chapter, we describe methods of modulating the expression of transgenes at the transcriptional level. Using cAMP-response element-binding protein (CREB)-dependent promoters as examples, a tool for the precise tuning of gene expression by using different core promoters and by varying the binding affinity of transcription factor operator sites is described.
合成生物学,即工程学与生物学的融合,近年来迅速发展成熟,极大地增加了人工基因电路的复杂性。在哺乳动物细胞中部署复杂的合成基因电路需要建立对转基因表达非常精确且正交的控制。在本章中,我们描述了在转录水平调节转基因表达的方法。以依赖于环磷酸腺苷反应元件结合蛋白(CREB)的启动子为例,描述了一种通过使用不同的核心启动子以及改变转录因子操纵位点的结合亲和力来精确调节基因表达的工具。