HTGAA 2026: Individual Final Project Documentation SECTION 1: ABSTRACT My project aims to engineer an autonomous, glowing plant using the firefly light-producing system. If successful, it will be the first demonstration of an autonomously glowing plant host through the firefly-based pathway. The significance of this work lies in several values, including cross-kingdom metabolic engineering and the longstanding challenge of solving the missing steps in the luciferin bioluminescence pathway. More broadly, this project asks the question of whether a plant can be rewired to support sustained luciferase-based bioluminescence.
HTGAA 2026: Individual Final Project Documentation
SECTION 1: ABSTRACT
My project aims to engineer an autonomous, glowing plant using the firefly light-producing system. If successful, it will be the first demonstration of an autonomously glowing plant host through the firefly-based pathway. The significance of this work lies in several values, including cross-kingdom metabolic engineering and the longstanding challenge of solving the missing steps in the luciferin bioluminescence pathway. More broadly, this project asks the question of whether a plant can be rewired to support sustained luciferase-based bioluminescence.
The object is to develop a proof of concept for autonomous, glowing plants. The central hypothesis is that if plants are engineered to produce key luciferin precursors and convert those precursors into luciferin, along with expressing that luciferase in proper cellular compartments, the plant will be capable of generating light autonomously.
To test this, the project will pursue several steps. First, validate transformation and expression workflows. Second, assemble and introduce a multi-gene pathway. Third, assess whether transformed plants produce measurable bioluminescence.
My project includes molecular cloning, Agrobacterium-mediated plant transformation, transient expression assays and imaging based detection of luminescence. Expected outcomes include validated plant expression, a pathway prototype, and evidence evaluating whether autonomous bioluminescence in plants using fireflies is feasible. If successful, this could provide foundation for future work in biosensing, sustainable light, and synthetic biology education.
SECTION 2: PROJECT AIMS
Aim 1: Experimental Aim
The first aim of my final project is to establish proof of concept for autonomous firefly bioluminescence in plants. This will be done by utilizing molecular cloning, Agrobacterium plant transformation, transient expression and luminescence imaging. This aim focuses on validating plant transformation, testing the core components needed, testing my experimental precursors and determining if the plant tissue can produce detectable bioluminescence.
Aim 2: Development Aim
The second aim of my project is to optimize the validated concept, and expand the bioluminescence pathway by improving availability, efficiency and light output. If Aim 1 is successful, this step would address technical limitations such as incomplete pathway flux and low brightness in order to develop a more robust bioluminescent plant system.
Aim 3: Visionary Aim
The third aim of my project is to develop this new plant pathway into a new platform, with applications expanding into biosensing, lighting, and education. If successful, this work could validate cross-kingdom metabolic pathways, while enabling a new way to visualize gene expression, metabolism and environmental responses in living organisms and other tests. Examples include non-invasive biosensing and reporting systems, use in cancer research for identifying tumours, non-toxic light use for festivals and fishing, and more.
SECTION 3: BACKGROUND
Background and Literature Context
Provide background research that explains the current state of knowledge and identifies the gap your project addresses.
Briefly summarize two peer-reviewed research citations relevant to your research (minimum four sentences).
Analysis of a Transcription Factor Using Transient Assay in Arabidopsis Protoplasts (Ow et al. (1986) demonstrated that the firefly luciferase gene can be expressed in transgenic plants and plant cells. This is an essential proof-of-concept paper that plants are able to produce light when given a luciferin substrate, establishing luciferase as a plausible reporter system. However, the plants in this experiment were not autonomous, and had to be sprayed with external luciferin to produce light. I will be building on this paper by adding genes that produce the luciferin precursors in the plant. However, this paper remains integral to my project as it proves the firefly light-producing enzyme can function in a plant host.
Plants with genetically encoded autoluminescence Mitiouchkina et al. (2020) demonstrated engineered tobacco plants using the fungal bioluminescence pathway. This experiment was the first in the world to produce autonomously glowing plants without external substrate. This paper is important to my work, as it proves a major advance in self-sustained plant luminescence as biologically feasible when metabolic pathways are compatible. However, their system was fungal, which is magnitudes less efficient than firefly luminescence. This paper is relevant as it provides a successful example, while emphasizing the novelty of attempting the same goal with a different bioluminescence system.
Explain how your project is novel or innovative (minimum three sentences).
My project is innovative as it would be the first to apply current knowledge of firefly bioluminescence to a challenge that has not been solved in plants yet: autonomous light production without external luciferin. We have been using firefly luciferase as reporter systems for a long time in biology, but have been constrained by its substrate being external, limiting it as a measurement tool. This project proposes a new system by reconstructing these missing metabolic steps, and completing the engineering needed to master one of the most efficient known biological light-producing systems.
This project is also novel as it combines cross-kingdom metabolic engineering, pathway design, and plant synthetic biology beyond transferring a single gene. There is coordination between precursor production, luciferin formation and enzyme localization all at the same time. This challenges the assumption that complex animal-associated bioluminescence pathways cannot be functional in plants, and if successful, could expand the boundaries of synthetic biology.
Explain why your project matters and what impact it could have (minimum five sentences).
This project matters because it addresses a major limitation in plant synthetic biology. Although bioluminescence systems are powerful tools in research, our dependence on external luciferin has limited the reporter system. Solving this pathway can not only advance our scientific knowledge of complex, multi-step pathways, but also expand technical capabilities by creating new self-reporting plants that do not require repeat substrate addition.Furthermore, improving our understanding of firefly bioluminescence could have broader relevance beyond plants, since firefly luciferase are widely used in biomedical research including imaging, gene expression studies, and experimental tracking applications. Beyond immediate research settings, autonomously growing plants could also support future applications in environmental monitoring, biosensing and synthetic biology education by providing simple, non-invasive reachout of activity. Broadly speaking, this project could help shift the field towards more ambitious plant engineering efforts, as plants can be designed not only to express individual genes, but carry out coordinated, functional traits.
Describe the ethical implications associated with your project and identify relevant ethical principles (minimum two paragraphs).
This project raises questions related to biosafety, environmental responsibility, and broader implications of engineering organisms for new traits. Although bioluminescence may be a harmless trait, it is still a genetically modified organism. Bio-safety is integral, where engineered organisms should not be released or handled irresponsibly. There should also be strong environmental responsibility, as synthetic traits produced in plants may have unintended consequences. The most relevant principles should be non-maleficence, where the research should not harm any ecosystems or public health, and responsibility, where the researchers involved address communication and long-term consequences. Lastly, the project should be transparent, creating useful scientific knowledge for the community.
To ensure ethical and societal responsibility, the project should be carried out under strict biosafety and containment practices. All genetically engineered plant material will be kept in controlled settings, and never released into the environment. Any experimental design will follow institutional guidelines for handling, storage and disposal so no unintended spread occurs. Transparency about uncertainty will be upheld, including the possibility the pathway fails, does not function as expected, imposes metabolic stress on the plant, or produces unintended effects. Responsible communication should also be upheld about any environmental questions behind it. The goal is to advance knowledge responsibly.
SECTION 4: EXPERIMENTAL DESIGN, TECHNIQUES, TOOLS, AND TECHNOLOGY
Create a detailed experimental plan for your final project. Include a timeline for each part (minimum 15 lines/sentences).
Include specific methods, tools, and technologies for each part of the project and analysis.
Describe expected results for each experiment.
Include figures if possible to show workflows.
Reminder: All HTGAA projects must include some DNA design!
Techniques Checklist
☐ Pipetting
☐ Lab Safety
☐ Bioproduction
☐ Registry of Standard Biological Parts
☐ Chassis Selection (e.g., DH5alpha)
☒ Bioethical Considerations
☐ Plasmid Preparation
☐ Bacterial Culturing
☐ Quality Control/Analysis
☐ Bacterial Processing (Centrifugation, Lysis, DNA Purification)