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Synthetic biology tools for engineering Goodwin oscillation in Trypanosoma brucei brucei

Borg, Yanika; Alsford, Sam; Pavlika, Vasos; Zaikin, Alexey; Nesbeth, Darren; (2022) Synthetic biology tools for engineering Goodwin oscillation in Trypanosoma brucei brucei. Heliyon , 8 (2) , Article e08891. 10.1016/j.heliyon.2022.e08891. Green open access

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Abstract

Kinetoplastid protozoa possess properties that are highly divergent from the mammalian, yeast and bacterial cells more commonly used in synthetic biology and represent a tantalisingly untapped source of bioengineering potential. Trypanosoma brucei brucei (T. b. brucei), an established model organism for studying the Kinetoplastida, is non-pathogenic to humans and provides an interesting test case for establishing synthetic biology in this phylogenetic class. To demonstrate further the tractability of Kinetoplastida to synthetic biology, we sought to construct and demonstrate a Goodwin oscillator, the simplest oscillatory gene network, in T. b. brucei for the first time. We report one completed iteration of the archetypal synthetic biology Design–Build–Test–Learn (DBTL) cycle; firstly, using Ab initio mathematical modelling of the behaviour a theoretical, oscillatory, trypanosomal synthetic gene network (SGN) to inform the design of a plasmid encoding that network. Once assembled, the plasmid was then used to generate a stable transfectant T. b. brucei cell line. To test the performance of the oscillatory SGN, a novel experimental setup was established to capture images of the fluorescent signal from motion-restricted live cells. Data captured were consistent with oscillatory behaviour of the SGN, with cellular fluorescence observed to oscillate with a period of 50 min, with varying amplitude and linear growth trend. This first DBTL cycle establishes a foundation for future cycles in which the SGN design and experimental monitoring setup can be further refined.

Type: Article
Title: Synthetic biology tools for engineering Goodwin oscillation in Trypanosoma brucei brucei
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.heliyon.2022.e08891
Publisher version: https://doi.org/10.1016/j.heliyon.2022.e08891
Language: English
Additional information: Copyright © 2022 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords: Synthetic biology, Trypanosoma brucei, Live cell imaging, Synthetic gene networks, Oscillation
UCL classification: UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL
URI: https://discovery.ucl.ac.uk/id/eprint/10142656
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