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A ratiometric dual-color luciferase reporter for fast characterization of transcriptional regulatory elements

View ORCID ProfileE. González-Grandío, View ORCID ProfileG. S. Demirer, W. Ma, View ORCID ProfileS.M. Brady, View ORCID ProfileM.P. Landry
doi: https://doi.org/10.1101/2021.05.26.443018
E. González-Grandío
1University of California Berkeley, Chemical and Biomolecular Engineering, Berkeley, CA, USA
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G. S. Demirer
2University of California Davis, Department of Plant Biology and Genome Center, Davis, CA, USA
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W. Ma
1University of California Berkeley, Chemical and Biomolecular Engineering, Berkeley, CA, USA
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S.M. Brady
2University of California Davis, Department of Plant Biology and Genome Center, Davis, CA, USA
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M.P. Landry
1University of California Berkeley, Chemical and Biomolecular Engineering, Berkeley, CA, USA
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  • For correspondence: landry@berkeley.edu
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Abstract

Plant synthetic biology requires precise characterization of genetic elements to construct complex genetic circuits that can improve plant traits or confer them new characteristics. Transcriptional reporter assays are essential to quantify the effect of gene expression regulator elements. Therefore, transcriptional reporter systems are a key tool in understanding control of gene expression in biology.

In this work we construct and characterize a dual-color luciferase ratiometric reporter system that possesses several advantages over currently used reporters. It is ratiometric, reducing variability and increasing consistency between experiments; it is fast, as both reporters can be measured at the same time in a single reaction, and it is cheaper to perform than current dual-luciferase reporter assays. We have validated our system quantifying the transcriptional capability of a panel of promoters and terminators commonly used in synthetic biology with a broad range of expression magnitudes, and in a biologically relevant system, nitrate response.

Competing Interest Statement

The authors have declared no competing interest.

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-NC-ND 4.0 International license.
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Posted May 27, 2021.
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A ratiometric dual-color luciferase reporter for fast characterization of transcriptional regulatory elements
E. González-Grandío, G. S. Demirer, W. Ma, S.M. Brady, M.P. Landry
bioRxiv 2021.05.26.443018; doi: https://doi.org/10.1101/2021.05.26.443018
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A ratiometric dual-color luciferase reporter for fast characterization of transcriptional regulatory elements
E. González-Grandío, G. S. Demirer, W. Ma, S.M. Brady, M.P. Landry
bioRxiv 2021.05.26.443018; doi: https://doi.org/10.1101/2021.05.26.443018

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