Anderson J Christopher, Clarke Elizabeth J, Arkin Adam P, Voigt Christopher A
Howard Hughes Medical Institute, California Institute of Quantitative Biology Department of Bioengineering, University of California, 717 Potter Street, Room 257 Berkeley, CA 94720, USA.
J Mol Biol. 2006 Jan 27;355(4):619-27. doi: 10.1016/j.jmb.2005.10.076. Epub 2005 Nov 14.
Bacteria can sense their environment, distinguish between cell types, and deliver proteins to eukaryotic cells. Here, we engineer the interaction between bacteria and cancer cells to depend on heterologous environmental signals. We have characterized invasin from Yersinia pseudotuburculosis as an output module that enables Escherichia coli to invade cancer-derived cells, including HeLa, HepG2, and U2OS lines. To environmentally restrict invasion, we placed this module under the control of heterologous sensors. With the Vibrio fischeri lux quorum sensing circuit, the hypoxia-responsive fdhF promoter, or the arabinose-inducible araBAD promoter, the bacteria invade cells at densities greater than 10(8)bacteria/ml, after growth in an anaerobic growth chamber or in the presence of 0.02% arabinose, respectively. In the process, we developed a technique to tune the linkage between a sensor and output gene using ribosome binding site libraries and genetic selection. This approach could be used to engineer bacteria to sense the microenvironment of a tumor and respond by invading cancerous cells and releasing a cytotoxic agent.
细菌能够感知其周围环境，区分细胞类型，并将蛋白质传递给真核细胞。在此，我们设计使细菌与癌细胞之间的相互作用取决于异源环境信号。我们已将假结核耶尔森菌的侵袭素鉴定为一种输出模块，该模块能使大肠杆菌侵入包括HeLa、HepG2和U2OS细胞系在内的癌细胞衍生细胞。为了在环境方面限制侵袭，我们将此模块置于异源传感器的控制之下。利用费氏弧菌的群体感应电路、缺氧反应性fdhF启动子或阿拉伯糖诱导型araBAD启动子，细菌分别在厌氧生长箱中生长后或在存在0.02%阿拉伯糖的情况下，以大于10⁸个细菌/毫升的密度侵入细胞。在此过程中，我们开发了一种利用核糖体结合位点文库和基因筛选来调节传感器与输出基因之间联系的技术。这种方法可用于设计细菌，使其感知肿瘤的微环境，并通过侵入癌细胞并释放细胞毒性剂来做出反应。
