An Interview with Dr Craig Martin on the Benefits of a New IVT Platform for RNA Manufacturing

Can you imagine a future where RNA therapeutics are routinely used as standard-of-care? A future where personalized medicine eases human suffering from diseases and disorders?  Dr. Craig Martin envisions this future with an eye on what advances are needed to get us there. To expedite discovery and expansion in healthcare, we need to solve one persistent challenge: scaling RNA manufacturing systems that deliver high purity efficiently and cost-effectively.

Dr. Craig Martin is the president and CSO of Waterfall Scientific™  and an Emeritus Professor in the Department of Chemistry at the University of Massachusetts Amherst. He has studied T7 RNA polymerase systems for over forty years. To help fulfill the promise of RNA therapeutics, Waterfall Scientific will commercialize the DuoTether™ technology, which was developed in his UMass lab. In the latest podcast from NEB’s series, Lessons from Lab & Life™, Dr. Martin explains the design of DuoTether technology and how it addresses the biggest challenges in RNA manufacturing today: purity, enzymatic synthesis of long RNAs and the overall cost to produce a product.

Solving pain points in RNA manufacturing with a novel IVT platform

For background, RNA manufacturing typically proceeds through several core stages: DNA template preparation, in vitro transcription (IVT), removal of the template and reaction components, purification, and finally quality analysis and formulation.

If you’re working in this space, you know that current RNA manufacturing introduces impurities that require downstream systems to clean up RNA, and none of this scales well. To put it simply, the post-IVT reaction components removal and purification steps are a real slog. These constraints are exactly what motivated Dr. Martin’s lab to rethink how IVT systems could be designed from the ground up.

Initial removal of template and IVT reaction components relies on chromatography methods such as Oligo (dT) and silica adsorption, and often, tangential flow filtration (TFF). These are expensive and difficult to scale. At this point, you have the desired RNA product, but you still have double-stranded RNA (dsRNA) byproducts from synthesis, self-complementary sequences, and truncated transcripts. RNA purification is crucial. Even trace amounts of dsRNA in RNA therapeutics can suppress translation or compromise tolerability in patients. At this stage, most commercial workflows use high-pressure liquid chromatography or specialized affinity steps to reduce the amount of double-stranded RNA. These are the most effective RNA purification methods. And yet again, they do not scale linearly. Instead, these methods are capacity- limited, expensive, and often sequence dependent.

Mind you, as you’re purifying, you’re losing product at each step.
 
In this podcast, Dr. Martin shares details on the design of DuoTether technology and the multiple improvements it enables, including the prospect of scaling out continuous, enzymatic flow reactions. If you’d like to review some data, the Martin lab’s recent report in Nucleic Acids Research demonstrates how the IVT platform can enable high yield, high purity mRNA synthesis with post-synthesis purification. It’s going to be exciting watching this technology translate into RNA therapeutics — particularly as the field seeks manufacturing platforms that are not only high performing, but truly scalable.