The Brains Behind the Next Trend in Biotech: Room-temperature Storage of Nucleic Acids

Lee Organick, PhD
12 min readJul 2, 2024

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As the cost of sequencing has continued to drop, nucleic acids (such as DNA and RNA) have given us more insights into our health. Now, nucleic acids are essential for scientific research and medical care. However, their storage poses a significant challenge. Currently, the vast majority of samples are frozen at -20°C or -80°C, a process that takes an enormous amount of electricity and leaves labs vulnerable to freezer or electricity failures.

Estimates show that the storage of DNA and RNA is expected to continue to increase over time. Figure adapted from Nova 1 Advisor.

An innovative movement is emerging to address these challenges: room-temperature storage of nucleic acids. This approach eliminates freezing, significantly reducing space and energy requirements. It also enhances the resilience of collections, protecting valuable samples from power outages and freezer failures.

We’ve delved into the minds behind the next big trend in biotech by interviewing experts from four pioneering organizations: Cache DNA, Imagene, 300K Solutions, and EnsiliTech. Discover their motivations, innovative methods, and visions for the future of room-temperature nucleic acid storage.

Question 1: What is your company’s method for storing nucleic acids at room temperature?

Lee Organick, Cache DNA

At Cache DNA, we encapsulate nucleic acids with a micro-scale process (known as “Caching”). During Caching, nucleic acids (negatively charged) are mixed with inert beads (positively charged), forming weak bonds. An encapsulant is mixed in, forming a protective shell around the nucleic acids and shielding the sample from degradants. This allows Cached samples to be kept at room temperature with prolonged degradation rates.

A magnified illustration of a single encapsulated bead (not to scale). Each one of these encapsulated beads is only a few microns in diameter. An animation of the Caching process can be found on Cache’s website.

When a Cached sample is ready to be retrieved, a total or partial aliquot is prepared and added to an etching solution. In 10 minutes, the solution dissolves both the encapsulant and bead core but does not react with the nucleic acids, leaving them free in solution for downstream applications. Importantly, no part of this process requires refrigeration, and the process is automatable and readily scalable.

Marthe Colotte, Imagene

Our innovation was based on the determination of the most stringent conditions for the optimal preservation of DNA and RNA at room temperature, which we have demonstrated to be indispensable for long-term storage and, in this case, for the preservation of DNA over thousands of years. The choice of a technological implementation of these conditions led us to invent the DNAshell and RNAshell capsules. They consist of a physical encapsulation that protects nucleic acid samples from atmospheric degradation factors (light, water, oxygen, ozone…). To achieve these rigorous conditions and to keep them airtight, these stainless-steel capsules are laser-welded under an inert gas. So, kept away from deleterious factors, nucleic acids can be transported and stored stand-alone for the very long term at true ambient conditions in a form compatible with any downstream use or analysis.

The Bioshielder encapsulation stations developed by Imagene enable containment, laser welding and leak testing of DNAshell and RNAshell capsules. Encapsulation is primarily service-based, but Imagene also sells industrial/ fully automated encapsulation stations with various throughputs.

Left: the DNA/RNAshell capsule. Right: the inside view of the Bioshielder Progress station. More information can be found on Imagene’s website.

Pablo Peñalosa Jiménez, 300k

300K Solutions has evolved freeze drying to the next level of precision. Several patented innovations allow high thermal homogeneity, precise pressure control, and the development of freeze-drying cycles tailored to each type of biological sample, allowing us to stabilize biological materials at room temperature. We have created a simple process where the user just needs to add the sample to our pre-coated vials, load the S3 device, and start the process without making any instrument setup.

An image of the 300k Solutions technology, which measures 134 x 62 x 77 cm. More information can be found on the 300k Solutions website.

300K Solutions makes freeze-drying accessible and easy to use. Our system also allows for broad flexibility since we can stabilize several biological materials, such as serum, plasma, solid tissues, and peripheral blood, that can all be used in research in the same manner as frozen samples.

Asel Sartbaeva, EnsiliTech

Ensilication is a chemical method for wrapping biopharmaceuticals in protective layers of silica on a molecular level to preserve the molecule better. To visualize how it works, a good analogy would be dipping a strawberry in dark chocolate and seeing it dry and harden around the fruit. One can repeat this several times and put on more layers of chocolate, which follows the strawberry shape, creating a hard shell that hardens around the strawberry. Like the chocolate, silica will grow around the target biomolecule in a bespoke, tailor-fitted manner, creating a “harder” shell that protects the biopharmaceutical product.

A representation of the ensilication process. More information can be found at EnsiliTech’s website. Image source.

This works because most biopharma products are made of amino-acid chains (proteins, viruses, enzymes, antibodies), which unfold and aggregate (stick to each other) at elevated temperatures, rendering them useless. We recently started looking at newer, mRNA-based vaccines, which today require ultra-cold temperatures down to -80°C. Being able to store and transport those at elevated temperatures, especially room temperature, would mean less wastage, less CO2 outputs from the cold-chain, and hopefully more people vaccinated in time to prevent many infectious diseases.

Question 2: How did you get into the room temperature storage field?

Lee Organick, Cache DNA

I began working in a biological research lab in 2012 and have had plenty of first-hand experience with freezers since then. I know on a visceral level how annoying they are to work with, maintain, and keep organized. Over time, my research shifted from generating and managing samples in a freezer to evaluating different preservation methods to store DNA long-term at room temperature (Organick et al. 2021). As a biologist and avid outdoorswoman, I needed to find a company passionate about making the world a better place. I was excited to join and couple my previous experience in the nucleic acid preservation area with the company’s mission to develop methods to store biomolecules more sustainably.

Marthe Colotte, Imagene

As a teenager, after a brief phase when I thought about becoming a forensic pathologist, I discovered forensic science, even though it was not yet popular. At the end of my engineering studies, I landed an internship at Imagene and stayed there because its founders instilled in me their passion for DNA stabilization. I found it incredibly rewarding to join a start-up in its early stages, to do my PhD there, and to be able to play an active part in its development. Even though we’ve had our ups and downs, it’s been an exciting adventure. While Imagene happened to be ahead of its time initially, now we have over 15 years of hindsight on the stability of our encapsulated samples, just when the biobank market is starting to consider ambient storage seriously.

Pablo Peñalosa Jiménez, 300k

I have been linked to the world of clinical diagnostics for the last 15 years, which has allowed me to be in contact with researchers and those responsible for carrying out clinical studies. During that time, I saw that ultra-low-temperature freezers (ULT freezers) were the preservation method of choice in the industry, but I was not satisfied. Four years ago, we conducted a feasibility study and showed that these ULT freezers cannot remain the industry’s top choice. They require a lot of space on the laboratory floor, which is very expensive, and they need temperature and electrical control systems to avoid mechanical failures that put the sample at serious risk of loss. Furthermore, transporting the samples while maintaining very low temperatures is problematic and increases the risk of sample loss previously mentioned. What drew me personally to join my colleagues at this company was our commitment to innovation and our mission to make a positive impact. I was inspired by the opportunity to be part of a team pushing boundaries and creating meaningful change in the field of room temperature biological sample handling.

Asel Sartbaeva, EnsiliTech

I took my daughter to be vaccinated in 2010 when she was just five days old. I saw that the doctor removed the vaccine from the fridge, so I asked if they should wait for the vaccine to be warmed to room temperature. The doctor explained that vaccines stop working if stored outside the fridge temperatures. This sparked my interest, so I researched why vaccines spoil outside the fridge. That’s when I learned that even now, about a quarter of vaccines spoil due to cold-chain failures, and 1.5 million infants worldwide die every year from vaccine-preventable diseases. This motivated me to start researching vaccine thermal stabilization. My background was in silica-based materials, so I looked at how silica could be applied for vaccine preservation. In 2017, a colleague, Dr. Stephen Wells, and I filed the first patent for vaccine thermal stabilization, called ensilication, which we invented.

Question 3: People have many reasons to consider using ambient storage. Can you share some examples you’ve heard?

Lee Organick, Cache DNA

One person I talked to at a biobanking conference had half of their freezers die in the same week. Since then, they’ve been traumatized and are much more eager to try room-temperature solutions that don’t rely on freezers.

In my research experience, I once worked in a lab with two -80°C freezers. It was Friday at 4:30 p.m. on a 4th of July weekend when one of the freezers died. Nobody was around to help. We lost a lot of samples that day, and I consider that the experience that planted the seed in me to move away from freezers!

But I’ve also talked to folks at large institutions, and because everything gets logistically easier when freezers aren’t involved, the implementation details alone are enough to make them eager for alternatives.

Marthe Colotte, Imagene

Anyone who works for or with laboratories has experienced (or at least knows of) tragic stories of losing precious samples due to a freezer failure or liquid nitrogen shortage. We’ve also had customers who were fed up with deciding what to do with their samples because they had no room for a new freezer in their facility’s basements. No matter how well-resourced they are or where they’re from, all these professionals dream of getting rid of the mental (and financial) burden of the cold chain.

Pablo Peñalosa Jiménez, 300k

Once you start working in biorepositories, you quickly hear about incidents frequently occurring when ultra-low freezers break down. For instance, my labmates told us that when they worked in repositories, they would take turns being on call and moving the samples in case of an alarm. They also say it was common to go to the lab at 2 a.m. or even over the weekend in an emergency. Therefore, there are several reasons for moving to room temperature sample stabilization.

Asel Sartbaeva, EnsiliTech

I heard from a WHO employee in a low-income country that once, a shipment of vaccines had arrived in the country and was waiting to clear customs. Sadly, the process was so long and challenging that it took them two weeks to get the shipment through. All vaccines were sitting on the tarmac for those two weeks in the country where summer temperatures can rise to over 35°C. After looking at the logged temperature data, they had to throw away almost 90% of the shipment, which showed that cold-chain was broken.

In another story, during the height of the COVID-19 pandemic, a shipment of COVID-19 vaccines arrived in a country and was stored for a couple of days in a facility arranged by the Ministry of Health. When they wanted to distribute the vaccines, they noticed one freezer was off. An investigation found that the cleaner had unplugged the freezer to charge their phone in the evening and forgot to switch it on again. All vaccines from this freezer were deemed unusable.

Question 4: Ambient preservation techniques aren’t new (spotting samples onto paper to be held at room temperature has been used since the 1960s!). From your perspective, what has been the greatest challenge to widespread adoption?

Lee Organick, Cache DNA

The biggest hurdle is changing labs’ entrenched habits. Transitioning from familiar freezing methods to new room temperature techniques can be daunting, especially for critical long-term storage. It’s so easy for labs to keep doing a process they’re used to. And with something as critical as storing RNA/DNA, I completely understand the caution of switching to a different method/protocol. Comprehensive educational materials on preservation science are essential to facilitate this shift.

Marthe Colotte, Imagene

On the one hand, habits die hard. People are reluctant to convert their valuable samples to a room temperature format even when presented with eloquent data. On the other hand, some stories circulate involving sub-optimal ambient preservation solutions that were adopted without sufficient validation. Finally, there are generally different entities within an organization that independently bear the energy costs, buy the freezers, and even manage the biobank. This often makes switching to ambient storage solutions a complex, coordinated effort across multiple leaders in an organization. However, in recent years, we have seen a professionalization of practices, making it much easier for biobanks to realize the benefits of room temperature storage.

Pablo Peñalosa Jiménez, 300k

When a new method changes the paradigm, there is always a reluctance to change unless we prove that the alternatives we present are as good as current methods. From this perspective, at 300K Solutions we are collecting data from our internal studies and establishing collaborations with real users at external laboratories to show that the performance of biological samples stabilized at room temperature is comparable with ultra-low temperature methods.

Asel Sartbaeva, EnsiliTech

Within the vaccine community, several obstacles and challenges need to be overcome before ambient preservation techniques can be adopted. For decades, vaccines were mainly developed with efficacy and safety in mind, and temperature stability was not considered important. Regulatory approvals are quite challenging as well, in addition to being costly and lengthy, making the developers and manufacturers focus mainly on efficacy and safety.

Question 5: What do you hope for the future of biosample storage? What might change?

Lee Organick, Cache DNA

If these preservation advances continue to happen at room temperature, as I think they will, I hope this technology will ultimately help patients on two fronts:

1. I hope advancements in room temperature storage will enhance patient access to medical testing through decreased operational costs. We have interfaced with several groups in the United States that can only send out samples infrequently because shipping frozen/refrigerated samples is too expensive.

2. Ultimately, I hope that room-temperature preservation science will enable samples to be stored longer so that the same sample can be analyzed 10, 20, or 50 years later with new and emerging technologies that didn’t exist when the sample was collected, helping unlock the mysteries behind rare diseases and other critical questions.

Marthe Colotte, Imagene

We hope for global awareness of climate change’s acceleration. However, regulations and policies to reduce greenhouse gas and carbon emissions may speed up biobanking managers’ adoption of new methods to store and transport samples at room temperature.

We also expect innovation and stimulation in the field of DNA storage with the emergence of a new application: storing digital data in DNA. I believe that, by storing large quantities of synthetic DNA for the DNA data storage field, it will become more acceptable to store biological samples at room temperature (using high-performance, validated technologies!).

Pablo Peñalosa Jiménez, 300k

Maintaining cold chains is a significant hurdle for research and clinical trials. There are large sectors of the global population where maintaining the necessary conditions to have a biorepository is extremely complicated. These small changes to start using room temperature technologies can help make biobanking much less complicated, and ultimately, while providing alternatives to the cold-chain may help richer countries to be more efficient, we hope it will allow those with lower resources to actively participate in the global research community.

Asel Sartbaeva, EnsiliTech

EnsiliTech’s mission is to bring vaccines to everyone around the globe and protect as many people as possible from preventable disease and death. Public awareness of the vaccine manufacturer and cold-chain logistics rose during the COVID pandemic and has brought the issues of vaccine thermal stability to the forefront. On the broader horizon, manufacturers are now signing new ECG promises, advocating CO2 reductions. This means they will inevitably turn to greener processes: new technologies that allow change for greener production, storage, and transport. As ensilication and many other new technologies are greener, the future looks brighter!

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