Integrated Human Practices
To better understand the real-world context of our project, we connected with several experts and stakeholders. Their invaluable insights have directly shaped our research, design, and implementation strategies.
Katalin Karikó, PhD
Nobel Laureate, Senior Vice President, BioNTech, and Adjunct Professor of Neurosurgery, University of Pennsylvania
To gain further context on the field of mRNA vaccine development during the COVID-19 pandemic, we reached out to the biochemist Dr. Katalin Karikó. She was awarded the 2023 Nobel Prize in Medicine for her discovery along with Dr. Drew Weissman that enabled the modified mRNA technology used in Pfizer-BioNTech and Moderna’s vaccines to prevent COVID-19 infection.
In response to our question about the biggest challenges faced when introducing novel vaccine technologies to the public, she highlighted an issue we anticipate with incorporating tardigrade IDPs into vaccine formulations. For mRNA-LNP vaccines, a major challenge is the chemical interaction between the mRNA and the ionizable lipid. This interaction can form adducts that render the mRNA untranslatable. She also provided two publications that further detail this problem.
Packer, M., Gyawali, D., Yerabolu, R. et al. A novel mechanism for the loss of mRNA activity in lipid nanoparticle delivery systems. Nat Commun 12, 6777 (2021). https://doi.org/10.1038/s41467-021-26926-0
Peronin, S., Malburet, C., Daher-Hassan, C., Picard, C., Willemin, T., Costamagna, F., Even, L., Eynard, T., Bensaid, F., Fertier-Prizzon, S., & Francois-Heude, M. (2025). Analytical approach for identification and mechanistic insights into mRNA-lipid adduct formation. Molecular therapy. Nucleic acids, 36(3), 102684. https://doi.org/10.1016/j.omtn.2025.102684
Marlo Walker, BSN, RN
Director of Health Services, University of Texas at Dallas Student Health Center
Our research directly focuses on a growing problem in the Dallas community and broader North Texas region of intense climate change and bipolar temperatures. This uncertainty of cascading effects was most prevalent during the 2021 snowstorm that knocked out a majority of power in Texas, with little to no backup power sources being separate from the national grid. Engaging with the UT Dallas Health Clinic focused on understanding challenges in vaccine storage, transportation, and administration in the context of extreme weather such as the 2021 Texas snowstorm.
We spoke with Marlo Walker, the director of health services with 15 years of experience working at the health center, overseeing logistics and primary and preventative care services to enrolled students. The health clinic serves about 200-300 students per month. Since the start of the pandemic, the clinic has administered 710 Moderna COVID-19 mRNA vaccines since the start of the pandemic. Due to the high cost of ultra-cold storage, the clinic opted for Moderna’s COVID-19 mRNA vaccines to store at -20°C over Pfizer’s COVID-19 mRNA vaccines that require storage at -80°C.
During the 2021 snowstorm, 1,370 vaccines were compromised due to a power outage as backup generators for the pharmacy and freezer failed to activate. The affected vaccines included nine different vaccines from four manufacturers, all of which had to be replaced, resulting in an estimated loss of $20,000. While backup generators are in place, they are often unreliable. This prompted the UTD Health Center to explore alternative methods and locations for safely storing vaccines during emergencies.
The health clinic also holds vaccine clinics at multiple locations across campus. Vaccines are transported across campus in a compact refrigerator maintained between 2-8°C, with temperatures monitored hourly days leading up to and during the clinic to ensure viability. Additionally, styrofoam coolers with stacked ice packs and aluminum insulation are used, with garden thermometers constantly monitoring. Tight packing can occasionally cause temperature fluctuations, potentially affecting stability. However, limited storage space makes it a logistical challenge to balance available space with maintaining optimal temperatures. Thus, the clinic uses data logger probes to monitor temperatures, placing vaccines on different shelves since cooling varies within the refrigerator. To maintain stable temperatures, frozen water bottles are used alongside shelf placement.
Ms. Walker agreed our plan was feasible and addressed a real issue faced by UT Dallas students, particularly international students, who rely on the health center for essential services. Without proper vaccine storage during emergencies, these students could go without preventative health care. This insight reinforced our motivation to pursue the project in its early stages and inspired us to share it with the broader collegiate community at the 2025 UT Dallas SPUR Symposium, where many domestic and international students who rely on the health center will learn about our work.
2025 University of Texas at Dallas Summer Platform for Undergraduate Research (SPUR) Symposium
Community Engagement Event
We presented our preliminary data and results to the wider UT Dallas community at the 2025 UT Dallas Summer Platform for Undergraduate Research (SPUR) Symposium. During this event, we practiced presenting our pitch and received live feedback from a diverse audience, including members from the honors college, principal investigators from different labs, undergraduate and graduate researchers, and even friends and family members of researchers from various industries. Through this experience, we gained valuable insights such as strategies to improve protein purification using protease inhibitors, adjusting buffer systems to match the pH with the different isoelectric points of our proteins, and effectively explaining our project to those outside of science. We also received suggestions for introducing tardigrades in a way that engages non-scientific audiences.
During this symposium, we attended a presentation by Matthew Cadena regarding his research on Intrinsically Disordered Proteins (IDPs) from human Asf1 Isoforms. We were inspired to explore how his IDP purification approaches might align with ours and subsequently arranged a meeting with him.
SPUR PosterAdjusting buffer systems to match the pH with different isoelectric points of our tardigrade IDPs: We created two buffers, one with pH 6 and one with pH 8, respectively, to encompass our full range of IDPs with a vast range of isoelectric points
When introducing the project to other audiences in subsequent meetings, we focused on explaining our experimental process clearly, without delving too deeply into scientific principles, to make our information accessible to those in other non-scientific industries
Farra Kahalnik Cohen, MS, MPH
UT Southwestern HealthStreet Initiative
When considering the novelty of our research, we wanted to understand the perspectives of our local patient populations. During the COVID-19 pandemic, we observed how there was skepticism surrounding mRNA vaccines in our region due to their novelty and complex mechanisms. Thus, we sought to gain further insight by collaborating with UT Southwestern’s Office of Community Health and Research Engagement, serving North Texas’s diverse patient population. When speaking with Ms. Farra Kahalnik Cohen, the manager of the aforementioned office, we learned that the office operates the HealthStreet initiative, a national program where Community Health Workers provide community members with free health screenings and health education. Through UT Southwestern’s involvement in this initiative, we were able to collate and analyze data from the Survey of Perceptions conducted as part of the Dallas HealthStreet hub.
This data helped us better understand the landscape of our project and the key factors we need to further consider for our development to move forward and reach patient populations. It also highlighted potential solutions of our project such as addressing access barriers commonly reported by patients and increasing transparency and trust with the community through our research outreach endeavors
University of Texas Southwestern Community Advisory Panel - Adult
Community Engagement Event
To build transparency and trust with community members, we connected with the UT Southwestern Community Advisory Panel (CAP). This panel provided us with direct perspectives from individuals across North Texas in different fields ,ranging from volunteers, health professionals, and leaders of non-profit organizations
During the CAP meeting, members recommended using clearer strategies to explain information to non-scientific audiences to ensure our science is accessible rather than “behind closed doors.” They resonated with our use of analogies to explain scientific concepts and encouraged this strategy. For example, we compared dog treats that help dogs take medicine to how mammalian cells more easily uptake plasmids to make complex concepts easier to understand. They also emphasized the use of animations and visuals, which encouraged us to continue utilizing this method in future presentation materials.
Although mRNA vaccines became more well-known during the COVID-19 pandemic, their mechanisms still remain not fully understood by the public. Members voiced their concerns about the safety of mRNA vaccines in the body, specifically questioning the dangers of injecting mRNA genetic material into the body and its degradation. They also wondered about the tardigrade protein itself and whether it is removed from the vaccine formulation before administration or if it remains in the vaccine until injection and must then be degraded by the body. These were important considerations we had not previously explored, which prompted us to conduct further research as we plan for the potential implementation and widespread public use of our project’s final product.
When asked about the intended audience for our research, our team was prompted to think more deeply as we had not yet fully defined our scope. This insightful question highlighted how we must tailor our educational materials and presentations for different levels of understanding. Through this discussion, we reflected on how current mRNA COVID-19 vaccine formulations such as Moderna, Pfizer, and Johnson & Johnson are designed for different age groups, vary in formulation, and require different storage conditions depending on the manufacturer.
From this, we concluded that our audience should be broad to include all age ranges and that feedback from diverse age groups would be essential. We also learned about the UT Southwestern Youth CAP and expressed interest in presenting our research to them to gain valuable perspectives from younger audiences.
University of Texas Southwestern Community Advisory Panel - Youth
Community Engagement Event
After deciding our population of focus would be individuals of all ages eligible to receive mRNA vaccines across different brands, we met with the UT Southwestern Youth Community Advisory Panel (Youth CAP). In this discussion, we asked members to share their experiences during the pandemic and how their own or their families’ health needs were affected.
For example, one member described how during the 2021 Texas snowstorm, their mother had to store her insulin vials outside in the snow to prevent them from thawing inside the house when the power went out. While this action risked the vial being damaged by natural debris, the danger of uncontrolled indoor temperatures was considered a greater threat to the medication’s potency. Another member recounted a similar situation with their grandmother’s medications, which also could not be properly temperature-controlled during the storm.
Such insights were valuable because they highlighted how natural disasters can destroy sensitive medications without proper storage, putting patients’ health at risk without a backup plan. The conversation made us consider how similar approaches like ours could be applied to other types of medications that require strict temperature control.
Jenny Molloy, PhD
Senior Research Associate and Shuttleworth Research Fellow at the University of Cambridge
She also is the Founder and Director of the Open Bioeconomy Lab and works to better understand problems facing researchers accessing biological research tools in low-resource contexts, particularly Latin America and Africa. She engages in policy work to examine the role and impact of open approaches to intellectual property and local manufacturing for the global bioeconomy.
She had previously seen tardigrades used as protectants in vivo and was enthusiastic to learn about our project.
When bearing more details about our project, she recommended that we investigate the amount of protein required to demonstrate measurable protection of mRNA. She also suggested that, in the long term, a stabilization period of four weeks at ambient temperature would be a valuable goal since it aligns with typical shipping timeframes. She clarified that this feedback was based on her knowledge of molecular reagents.
From a regulatory standpoint, she noted that because our project involves introducing modified vaccines into humans, it would eventually require regulatory review, as human research is highly regulated. However, she did not believe that our project posed any major biosafety risks that would demand excessive permissions. Additionally, she also noted that if our goal is to achieve vaccine equity, we must first determine whether cold chain limitations are truly the main bottleneck in vaccine distribution. If our project succeeds as a proof of concept, the next step would be to investigate other factors that influence equity and assess how our approach could address the broader challenges in vaccine access.
On a larger scale, we are aware that our project should consider how tardigrade proteins would be sourced globally, given that tardigrades are widely distributed around the world. In response, Dr. Molloy brought to our attention the Nagoya Protocol, which governs the use of genetic resources across countries and addresses issues such as “profiting” from materials obtained abroad. While later we discovered the United States is not a part of the Nagoya Protocol, it is worth noting that for an innovation like ours with potential global impact, it is worthwhile to research and understand these considerations.
Regarding our His-tag system, Dr. Molloy explained that she typically uses a Strep-tag because of its high specificity but noted that it may not be compatible with our project. She agreed that a His-tag would be appropriate for our work as she has previously used it alongside size exclusion chromatography, a technique we spoke to Mr. Cadenas about previously. However, she mentioned that she has never worked with intrinsically disordered proteins and encouraged us to further investigate their properties.
The conversation made us consider the number of protein molecules required to protect a single molecule of DNA and recognized the importance of examining formulation ratios in mRNA vaccine preparations when designing our trial tests.She supported our decision to proceed with the His-tag purification system. We gained a global perspective on our project, particularly regarding manufacturing, worldwide distribution, and the implications of using biomaterials across borders through the Nagoya protocol.
Dan Barouch, MD, PhD
William Bosworth Castle Professor of Medicine
Nearing the end of our project, we were interested in discussing our project’s implications with other scientists who have created similar solutions to address cold-chain storage regarding vaccines. We came across Dr. Barouch’s research on microneedle vaccines being a delivery mechanism for thermostable COVID-19 mRNA vaccines to reach access to low-resource areas.
When discussing our experimental conditions, he emphasized the importance of making our trials as realistic as possible to the conditions vaccines face in the real world. As future directions, he suggested extending our stress test conditions for hours, days, or even months. Another condition included shipping samples to simulate real distribution challenges. During the testing stage, he advised us to prioritize temperature-specific conditions over freeze-thaw cycles, since mRNA vaccines are rarely taken in and out repeatedly in clinical or cold chain conditions. According to his expertise, the industry is aware of the vaccines’ delicate nature and avoids constant handling.
He agreed with our proof-of-concept plan to use the BCA assay as a measurable output. He emphasized making sure the protein-mRNA complex protects the mRNA from degradation without interfering with its function or reducing its immunogenicity.
Some future directions he emphasized include conducting stress test trials using mRNA-lipid nanoparticle (LNP) conjugates, since this combination represents the more accurate, realistic delivery system used in clinical trials versus mRNA alone without LNPs. He also suggested investigating whether IDPs provide more stability than current methods of vaccine stabilization such as the sugar trehalose or lipid nanoparticles themselves. In the long term, he questioned whether tardigrades would need to be removed from vaccine formulations before administration in humans. This concern was shared by the UT Southwestern Community Advisory Panel Adult group, and remains an important consideration for future applications.
Vander Straeten, A., Sarmadi, M., Daristotle, J. L., Kanelli, M., Tostanoski, L. H., Collins, J., Pardeshi, A., Han, J., Varshney, D., Eshaghi, B., Garcia, J., Forster, T. A., Li, G., Menon, N., Pyon, S. L., Zhang, L., Jacob-Dolan, C., Powers, O. C., Hall, K., Alsaiari, S. K., … Jaklenec, A. (2024). A microneedle vaccine printer for thermostable COVID-19 mRNA vaccines. Nature biotechnology, 42(3), 510–517. https://doi.org/10.1038/s41587-023-01774-z
Dr. Jeffery Way, PhD
Lecturer in the Department of Systems Biology and in the Laboratory of Systems Pharmacology at Harvard Medical Schoo
He had previously worked with the Secretory-Abundant Heat Soluble (SAHS) Tardigrade IDP, and provided us with his published literature on the topic.
Lim, S., Reilly, C. B., Barghouti, Z., Marelli, B., Way, J. C., & Silver, P. A. (2024). Tardigrade secretory proteins protect biological structures from desiccation. Communications biology, 7(1), 633. https://doi.org/10.1038/s42003-024-06336-w
Giovanni Traverso, MD, PhD
Assistant Professor of Medicine and Associate Physician in Division of Gastroenterology, Brigham and Women's Hospital (BWH)
He researched protecting healthy tissue from radiation damage using nanoparticle-delivered mRNA that encodes a damage-suppressor protein (DSUP) found in tardigrades.
When asking him what functional assays he recommended for assessing the protective efficacy of IDPs on nucleic acids in vitro, he mentioned referring to his publication where he utilized comet assays of cells pretreated with IDPs or control mRNA and exposed to stressors and fluorescence images.
Kirtane, A.R., Bi, J., Rajesh, N.U. et al. Radioprotection of healthy tissue via nanoparticle-delivered mRNA encoding for a damage-suppressor protein found in tardigrades. Nat. Biomed. Eng 9, 1240–1253 (2025). https://doi.org/10.1038/s41551-025-01360-5
Michael Cory, PhD
Synthetic Biology HIVE Fellow at Harvard Medical School, Silver Lab
When discussing our project’s goal of protecting mRNA vaccines in clinical settings, he mentioned that if we want to capture the same conditions that are likely to occur at clinics, refreezing our samples to test resilience is more appropriate compared to using liquid nitrogen baths or rapid-freeze equipment that clinics don’t have. He agreed that using freeze-thaw cycles is a reasonable way to simulate real-world clinic environments.
He asked whether we planned to use mass spectrometry or another method to confirm that the post-translational modifications match those found naturally. Our current plan was utilizing CHO and HEK cell lines as they have been researched to have similar post-translational modifications to human cells. Additionally, we don’t have access to more specific methods for double confirmation.
A key challenge he pointed out is that unstructured regions of IDPs make them susceptible to degradation by proteases, as the cell may recognize them as misfolded proteins. Large IDPs are difficult to optimize for expression because they do not fold properly. Additionally, he mentioned affinity tags may not work effectively because they can become masked by other regions of the protein that fold back on themselves. This aligned with our experience of utilizing His-tags, as they were not very specific, and we had been struggling to isolate our proteins. We then shared the challenges we were experiencing with IDP purification. He suggested some strategies, such as diluting or adjusting the salinity of the buffer system.
When explaining the nature of tardigrade IDPs, he mentioned IDPs may tend to form phase-separated condensates. Based on his experience with protein aggregates, he mentioned there is an unrecoverable type and another that is naturally prone to aggregation but can be coaxed back into solution.
We maintained our freeze-thaw cycle conditions for the mRNA-IDP stress tests and added multiple cycles to simulate the clinical environment where vaccines may be repeatedly taken in and out of cold storage.
Matthew Cadena
Undergraduate Neuroscience Student at UT Dallas
Matthew Cadena researched Human Asf1 Isoforms, focusing on their C-terminal intrinsically disordered regions within the UT Dallas D’Arcy lab. We spoke with Matthew regarding his purification strategies for Asf1 IDPs, and he provided us with several valuable suggestions. These included adding a double dose of protease inhibitors and conducting all procedures at cold temperatures to preserve protein integrity. His experimental process also used size-exclusion and ion-exchange chromatography using proteins tagged with a Flag tag. He recommended analyzing our tardigrade gene’s disordered regions to learn more about their properties, such as charge, acidity/basicity, and whether the disordered regions are localized on the tail or distributed throughout based on their amino acid sequence.
This reinforced our view from SPUR that protease inhibitors are essential, leading us to include them in the purification process in the form of a cocktail tablet. It also confirmed that conducting our purification steps in the cold room is a best practice and to continue to do so. The conversation inspired us to explore online models of our proteins and look at protein interactions using AlphaFold.