Rapid prototyping and design of cybergenetic single-cell controllers

Sant Kumar, Marc Rullan and Mustafa Khammash ()
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Sant Kumar: ETH Zürich
Marc Rullan: ETH Zürich
Mustafa Khammash: ETH Zürich

Nature Communications, 2021, vol. 12, issue 1, 1-13

Abstract: Abstract The design and implementation of synthetic circuits that operate robustly in the cellular context is fundamental for the advancement of synthetic biology. However, their practical implementation presents challenges due to low predictability of synthetic circuit design and time-intensive troubleshooting. Here, we present the Cyberloop, a testing framework to accelerate the design process and implementation of biomolecular controllers. Cellular fluorescence measurements are sent in real-time to a computer simulating candidate stochastic controllers, which in turn compute the control inputs and feed them back to the controlled cells via light stimulation. Applying this framework to yeast cells engineered with optogenetic tools, we examine and characterize different biomolecular controllers, test the impact of non-ideal circuit behaviors such as dilution on their operation, and qualitatively demonstrate improvements in controller function with certain network modifications. From this analysis, we derive conditions for desirable biomolecular controller performance, thereby avoiding pitfalls during its biological implementation.

Date: 2021
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DOI: 10.1038/s41467-021-25754-6

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