ACS Publications. Most Trusted. Most Cited. Most Read
Genetic Toggle Switch in Plants

OR SEARCH CITATIONS

My Activity
Publications

Figure 1Loading Img
Download Hi-Res ImageDownload to MS-PowerPointCite This:ACS Synth. Biol. 2025, 14, 6, 1988-2001
    Research Article

    Genetic Toggle Switch in Plants
    Click to copy article linkArticle link copied!

    • Tessema K. Kassaw
      Tessema K. Kassaw
      Department of Biology, Colorado State University, Fort Collins, Colorado 80523, United States
      More by Tessema K. Kassaw
    • Wenlong Xu
      Wenlong Xu
      Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado 80523, United States
      More by Wenlong Xu
    • Christopher S. Zalewski
      Christopher S. Zalewski
      Department of Biology, Colorado State University, Fort Collins, Colorado 80523, United States
      More by Christopher S. Zalewski
    • Katherine Kiwimagi
      Katherine Kiwimagi
      Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado 80523, United States
      More by Katherine Kiwimagi
    • Ron Weiss
      Ron Weiss
      Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States
      More by Ron Weiss
    • Mauricio S. Antunes
      Mauricio S. Antunes
      Department of Biology, Colorado State University, Fort Collins, Colorado 80523, United States
      More by Mauricio S. Antunes
      Orcidhttps://orcid.org/0000-0002-5453-1531
    • Ashok Prasad*
      Ashok Prasad
      Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado 80523, United States
      *Email: [email protected]. Phone: 970-491-5175.
      More by Ashok Prasad
      Orcidhttps://orcid.org/0000-0002-0832-8153
    • June I. Medford*
      June I. Medford
      Department of Biology, Colorado State University, Fort Collins, Colorado 80523, United States
      *Email: [email protected]. Phone: 970-491-7865.
      More by June I. Medford
      Orcidhttps://orcid.org/0000-0002-0599-4863
    Other Access OptionsSupporting Information (1)

    ACS Synthetic Biology

    Cite this: ACS Synth. Biol. 2025, 14, 6, 1988–2001
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acssynbio.4c00777
    Published May 19, 2025
    Copyright © 2025 American Chemical Society
    Request reuse permissions

    Abstract

    Click to copy section linkSection link copied!
    Abstract Image

    In synthetic biology, genetic components are assembled to make transcriptional units, and transcriptional units are assembled into circuits to perform specific and predictable functions of a genetic device. Genetic devices have been described in bacteria, mammalian cell cultures, and small organoids, yet the development of programmable genetic circuits for devices in plants has lagged. Programmable genetic devices require defining the component’s quantitative functions. Because plants have long life spans, studies often use transient analysis to define quantitative functions, while verification in stably engineered plants is often neglected and largely unknown. This raises the question if unique attributes of plants, such as environmental sensitivity, developmental plasticity, or alternation of generations, adversely impact predictability of plant genetic circuits and devices. Alternatively, it is also possible that genetic elements to produce predictable genetic devices for plants require rigorous characterization with detailed mathematical modeling. Here, we use plant genetic elements with quantitatively characterized transfer functions and developed in silico models to guide their assembly into a genetic device: a toggle switch or a mutually inhibitory gene-regulatory device. Our approach allows for computational selection of plant genetic components and iterative refinement of the circuit if the desired genetic functions are not initially achieved. We show that our computationally selected genetic circuit functions as predicted in stably engineered plants, including through tissue and organ differentiation. Developing abilities to produce predictable and programmable plant genetic devices opens the prospect of predictably engineering plant’s unique abilities in sustainable human and environmental systems.

    Copyright © 2025 American Chemical Society

    Subjects

    what are subjects

    Keywords

    what are keywords

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. Add or change your institution or let them know you’d like them to include access.

    Supporting Information

    Click to copy section linkSection link copied!

    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acssynbio.4c00777.

    • Schematic showing details of Toggle 1.0 genetic components; schematic showing details of Toggle 2.0 genetic components; luminescence heat maps (linear scale) of representative plants containing Toggle 1.0 under different treatment conditions; quantitative data analyses of Toggle 1.0 performance; numerical simulation results of the ODE model using parameter sets estimated by the MCMC method for Toggle 1.0; luminescence heat maps (linear scale) of representative plants containing Toggle 2.0 under different treatment conditions; quantitative data analyses of Toggle 2.0 performance; numerical simulation results of the ODE model using parameter sets estimated by the MCMC method for Toggle 2.0; luminescence heat maps (linear scale) of selected plants showing Toggle 2.1 behavior under different treatments; quantitative data analyses of Toggle 2.1 performance; numerical simulation results of the ODE model using parameter sets estimated by the MCMC method for Toggle 2.1; determination of chromosomal location and copy number estimation of the transgene; comparisons between dimensionless parameter values estimated from protoplast assays and whole plant luciferase data; sequence information on all the parts used to construct the toggle circuits; AD primers used for TAIL PCR; T-DNA left border primer sequences for pGREENII0229 vector; p-values of two-sample two-sided t-test between fold changes of different treatments of Toggle 1.0 shoots; p-values of two-sample two-sided t-test between fold changes of different treatments of Toggle 1.0 roots; p-values of two-sample two-sided t-test between fold changes of different treatments of Toggle 2.0 shoots; p-values of two-sample two-sided t-test between fold changes of different treatments of Toggle 2.0 roots; p-values of two-sample two-sided t-test between fold changes of different treatments of Toggle 2.1 shoots; p-values of two-sample two-sided t-test between fold changes of different treatments of Toggle 2.1 roots; parameter values used for the choice of Toggle 1.0 and Toggle 2.0 components; parameter values estimated from the protoplast assays using equation S4 with estimated Hill coefficient less than 6; designing principles of a bistable toggle switch; design of Toggle 1.0 and Toggle 2.0; data analysis of Toggle 1.0; MCMC parameter estimation results for Toggle 1.0; data analysis of Toggle 2.0; MCMC parameter estimation results for Toggle 2.0; data analysis of Toggle 2.1; MCMC parameter estimation results for Toggle 2.1; transgene insertion and copy number estimation; comparison of parameters estimated from protoplast assays and whole plant luminescence data; and proof of the comparability of dedimensionalized parameter values estimated from protoplast assays and whole plant luminescence data (PDF)

    Terms & Conditions

    Electronic Supporting Information files are available without a subscription to ACS Web Editions. The American Chemical Society holds a copyright ownership interest in any copyrightable Supporting Information. Files available from the ACS website may be downloaded for personal use only. Users are not otherwise permitted to reproduce, republish, redistribute, or sell any Supporting Information from the ACS website, either in whole or in part, in either machine-readable form or any other form without permission from the American Chemical Society. For permission to reproduce, republish and redistribute this material, requesters must process their own requests via the RightsLink permission system. Information about how to use the RightsLink permission system can be found at http://pubs.acs.org/page/copyright/permissions.html.

    Cited By

    Click to copy section linkSection link copied!

    This article has not yet been cited by other publications.

    Download PDF
    Go to ACS Synthetic Biology
    Get e-Alerts
    Get e-Alerts

    ACS Synthetic Biology

    Cite this: ACS Synth. Biol. 2025, 14, 6, 1988–2001
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acssynbio.4c00777
    Published May 19, 2025
    Copyright © 2025 American Chemical Society
    Request reuse permissions

    Article Views

    570

    Altmetric

    -

    Citations

    -
    Learn about these metrics

    Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

    Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

    The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.