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Genetically encoded, noise-tolerant, auxin biosensors in yeast facilitate metabolic engineering and directed evolution

Patarasuda Chaisupa, View ORCID ProfileMd Mahbubur Rahman, Sherry B. Hildreth, Saede Moseley, Chauncey Gatling, View ORCID ProfileRichard F. Helm, View ORCID ProfileR. Clay Wright
doi: https://doi.org/10.1101/2023.03.21.533585
Patarasuda Chaisupa
1Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, United States
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Md Mahbubur Rahman
1Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, United States
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Sherry B. Hildreth
2Fralin Life Sciences Institute, Virginia Tech, Blacksburg, VA 24061, United States
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Saede Moseley
1Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, United States
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Chauncey Gatling
1Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, United States
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Richard F. Helm
2Fralin Life Sciences Institute, Virginia Tech, Blacksburg, VA 24061, United States
3Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061, United States
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R. Clay Wright
1Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, United States
2Fralin Life Sciences Institute, Virginia Tech, Blacksburg, VA 24061, United States
4The Translational Plant Sciences Center (TPSC), Virginia Tech, Blacksburg, VA 24061, United States
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  • For correspondence: wrightrc@vt.edu
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Abstract

Auxins are important plant growth regulating compounds that are applied in vast quantities to crops across the globe to control weeds and improve crop quality and yield. Auxins are also produced by nearly every kingdom of life and control both organismal behavior as well as inter-kingdom interactions. Improving our understanding of auxin biosynthesis and signaling is critical to both improving crop plants and controlling symbiotic, commensal, and parasitic inter-kingdom relationships, many of which are critical to ecosystems from forests and oceans to the human microbiome. We present a suite of auxin biosensors that will advance our understanding of and ability to engineer auxin perception by plants and auxin production by fungi. We have developed genetically encoded, ratiometric auxin biosensors in the model yeast Saccharomyces cerevisiae, based on the mechanism plants use to perceive auxin. The ratiometric design of these biosensors improves measurements of auxin concentration by reducing clonal and growth phase variation. These biosensors are capable of measuring exogenous auxin in yeast cultures across five orders of magnitude, likely spanning the physiologically relevant range. We implement these biosensors to measure the production of auxin during different growth conditions and phases for S. cerevisiae. Finally, we demonstrate how these biosensors could be used to improve quantitative functional studies and directed evolution of plant auxin perception machinery. These genetically encoded auxin biosensors will enable future studies of auxin biosynthesis, transport, and signaling in a wide range of yeast species, as well as other fungi, and plants.

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Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/PlantSynBioLab/auxin-biosensor-data

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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-NC 4.0 International license.
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Posted March 21, 2023.
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Genetically encoded, noise-tolerant, auxin biosensors in yeast facilitate metabolic engineering and directed evolution
Patarasuda Chaisupa, Md Mahbubur Rahman, Sherry B. Hildreth, Saede Moseley, Chauncey Gatling, Richard F. Helm, R. Clay Wright
bioRxiv 2023.03.21.533585; doi: https://doi.org/10.1101/2023.03.21.533585
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Genetically encoded, noise-tolerant, auxin biosensors in yeast facilitate metabolic engineering and directed evolution
Patarasuda Chaisupa, Md Mahbubur Rahman, Sherry B. Hildreth, Saede Moseley, Chauncey Gatling, Richard F. Helm, R. Clay Wright
bioRxiv 2023.03.21.533585; doi: https://doi.org/10.1101/2023.03.21.533585

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