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Cheetah: a computational toolkit for cybergenetic control

View ORCID ProfileElisa Pedone, Irene de Cesare, View ORCID ProfileCriseida G. Zamora-Chimal, David Haener, Lorena Postiglione, Antonella La Regina, Barbara Shannon, View ORCID ProfileNigel J. Savery, View ORCID ProfileClaire S. Grierson, View ORCID ProfileMario di Bernardo, View ORCID ProfileThomas E. Gorochowski, View ORCID ProfileLucia Marucci
doi: https://doi.org/10.1101/2020.06.25.171751
Elisa Pedone
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
2School of Cellular and Molecular Medicine, University of Bristol, Biomedical Sciences Building, University Walk, BS8 1TD, Bristol, UK
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Irene de Cesare
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
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Criseida G. Zamora-Chimal
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
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David Haener
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
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Lorena Postiglione
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
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Antonella La Regina
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
2School of Cellular and Molecular Medicine, University of Bristol, Biomedical Sciences Building, University Walk, BS8 1TD, Bristol, UK
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Barbara Shannon
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
4School of Biochemistry, University of Bristol, Biomedical Sciences Building, University Walk, BS8 1TD, Bristol, UK
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Nigel J. Savery
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
4School of Biochemistry, University of Bristol, Biomedical Sciences Building, University Walk, BS8 1TD, Bristol, UK
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Claire S. Grierson
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
5School of Biological Sciences, University of Bristol, Tyndall Avenue, BS8 1TQ, Bristol, UK
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Mario di Bernardo
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
6Department of EE and ICT, University of Naples Federico II, Via Claudio 21, 80125, Naples, Italy
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Thomas E. Gorochowski
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
5School of Biological Sciences, University of Bristol, Tyndall Avenue, BS8 1TQ, Bristol, UK
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  • For correspondence: thomas.gorochowski@bristol.ac.uk lucia.marucci@bristol.ac.uk
Lucia Marucci
1Department of Engineering Mathematics, University of Bristol, Ada Lovelace Building, University Walk, BS8 1TW, Bristol, UK
2School of Cellular and Molecular Medicine, University of Bristol, Biomedical Sciences Building, University Walk, BS8 1TD, Bristol, UK
3BrisSynBio, Life Sciences Building, Tyndall Avenue, BS8 1TQ, Bristol, UK
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  • ORCID record for Lucia Marucci
  • For correspondence: thomas.gorochowski@bristol.ac.uk lucia.marucci@bristol.ac.uk
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Abstract

Advances in microscopy, microfluidics and optogenetics enable single-cell monitoring and environmental regulation and offer the means to control cellular phenotypes. The development of such systems is challenging and often results in bespoke setups that hinder reproducibility. To address this, we introduce Cheetah – a flexible computational toolkit that simplifies the integration of real-time microscopy analysis with algorithms for cellular control. Central to the platform is an image segmentation system based on the versatile U-Net convolutional neural network. This is supplemented with functionality to robustly count, characterise and control cells over time. We demonstrate Cheetah’s core capabilities by analysing long-term bacterial and mammalian cell growth and by dynamically controlling protein expression in mammalian cells. In all cases, Cheetah’s segmentation accuracy exceeds that of a commonly used thresholding-based method, allowing for more accurate control signals to be generated. Availability of this easy-to-use platform will make control engineering techniques more accessible and offer new ways to probe and manipulate living cells.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://www.github.com/BiocomputeLab/cheetah

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted April 19, 2021.
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Cheetah: a computational toolkit for cybergenetic control
Elisa Pedone, Irene de Cesare, Criseida G. Zamora-Chimal, David Haener, Lorena Postiglione, Antonella La Regina, Barbara Shannon, Nigel J. Savery, Claire S. Grierson, Mario di Bernardo, Thomas E. Gorochowski, Lucia Marucci
bioRxiv 2020.06.25.171751; doi: https://doi.org/10.1101/2020.06.25.171751
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Cheetah: a computational toolkit for cybergenetic control
Elisa Pedone, Irene de Cesare, Criseida G. Zamora-Chimal, David Haener, Lorena Postiglione, Antonella La Regina, Barbara Shannon, Nigel J. Savery, Claire S. Grierson, Mario di Bernardo, Thomas E. Gorochowski, Lucia Marucci
bioRxiv 2020.06.25.171751; doi: https://doi.org/10.1101/2020.06.25.171751

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