Programmable cell killing by rewiring MAPK signalling

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Kentaro Furukawa, Annika Volkwein, Paula Aguilera, Takako Furukawa, Stefan Hohmann

University of Gothenburg, Sweden

Engineered biological systems that execute precisely regulated responses are useful in a variety of applications. For instance, gene- or cell-based therapies for killing pathogenic cells can be engineered instead of development of novel drugs. However, there have been only a few proof of such concept so far. Here, we show approaches for specific input-triggered programmed killing of engineered or non-engineered yeast cells by rewiring the osmoregulatory high-osmolarity glycerol (HOG) MAPK pathway. Combinatorial integration of rich genetic parts such as a heterologous kinase, constitutively active components and chemical-inducible promoters allows controlling yeast growth via improper activation of the HOG pathway in a Boolean logic manner. Rewiring two upstream osmosensing branches of the HOG pathway as different logic functions (AND, NOR, or N-IMPLIES) enables also complex XNOR and XOR logic killing. Moreover, we propose a genetic-based predator-prey system, in which engineered cells kill specific target cells in a target cell-dependent manner. As a proof of concept, we demonstrate that engineered yeast cells (predator) carrying sleeping MAPK-disorder genes kill virtual pathogenic yeast cells (prey) in an AND logic manner. Thus, rewiring MAPK signalling with existing genetic parts provides new synthetic biological strategies useful for future medical applications.