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A Ratiometric Dual Color Luciferase Reporter for Fast Characterization of Transcriptional Regulatory Elements in Plants

González-Grandío, Eduardo and Demirer, Gozde S. and Ma, Wenhe and Brady, Siobhan and Landry, Markita P. (2021) A Ratiometric Dual Color Luciferase Reporter for Fast Characterization of Transcriptional Regulatory Elements in Plants. ACS Synthetic Biology, 10 (10). pp. 2763-2766. ISSN 2161-5063. doi:10.1021/acssynbio.1c00248.

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Plant synthetic biology requires precise characterization of genetic elements to construct complex genetic circuits that can improve plant traits or confer them with new characteristics. Transcriptional reporter assays are essential to quantify the effect of gene expression regulator elements. Additionally, 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, thus 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 less expensive 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.

Item Type:Article
Related URLs:
URLURL TypeDescription
González-Grandío, Eduardo0000-0002-2059-4542
Demirer, Gozde S.0000-0002-3007-1489
Brady, Siobhan0000-0001-9424-8055
Landry, Markita P.0000-0002-5832-8522
Additional Information:© 2021 American Chemical Society. Received 18 June 2021. Published online 14 September 2021. Published in issue 15 October 2021. M.P.L. acknowledges support of a USDA BBT EAGER award (2019-67013-29104) and a USDA NIFA award (2018-67021-27964). G.S.D. is supported by the Resnick Sustainability Institute Postdoctoral Fellowship, and G.S.D. and S.M.B. acknowledge support of a USDA BBT-EAGER award (2019-67013-29012), and partial support by an HHMI Faculty Scholar fellowship. pRedF was a gift from Koen Venken (Addgene #118057). Author Contributions. E.G. and M.P.L. conceived the project and wrote the manuscript. E.G., G.S.D., S.M.B., and M.P.L. designed the experiments. E.G. performed the experiments and data analysis. E.G. and W.M. prepared the plasmids. G.S.D. performed the nitrate experiments. All authors have edited and commented on the manuscript and have given their approval of the final version. The authors declare no competing financial interest.
Group:Resnick Sustainability Institute
Funding AgencyGrant Number
Department of Agriculture2019-67013-29104
Department of Agriculture2018-67021-27964
Resnick Sustainability InstituteUNSPECIFIED
Department of Agriculture2019-67013-29012
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Subject Keywords:synthetic biology transcriptional regulation reporter assay luciferase plant
Issue or Number:10
Record Number:CaltechAUTHORS:20220302-323877000
Persistent URL:
Official Citation:A Ratiometric Dual Color Luciferase Reporter for Fast Characterization of Transcriptional Regulatory Elements in Plants Eduardo González-Grandío, Gozde S. Demirer, Wenhe Ma, Siobhan Brady, and Markita P. Landry ACS Synthetic Biology 2021 10 (10), 2763-2766 DOI: 10.1021/acssynbio.1c00248
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113700
Deposited By: George Porter
Deposited On:03 Mar 2022 17:27
Last Modified:03 Mar 2022 17:27

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