Project Summary/Abstract
Since the birth of biotechnology in the 1980s, efficient nucleic acid synthesis has been a key
driver of biological discovery and bio-product development, culminating in the recent emergence of
the ‘synthetic biology’ field and rapid development of nucleic acid therapeutics. Sequence-specific
RNA and DNA manufacturing is having an especially strong impact in healthcare, where FDA
approval of the first oligonucleotide-based therapeutics in 2018 quickly led to many hundreds of
similar drugs entering clinical trials. Despite these positive developments, new DNA and RNA
synthesis technologies are urgently needed to meet the rapidly rising demand for clinical-grade
oligonucleotides, because the current chemical synthesis strategies are costly, difficult to scale,
inefficient and limited to molecules of 200 nucleotides or less in length. In the past decade, enzymatic
oligonucleotide synthesis (EOS) strategies have been explored and developed for this purpose.
In a Phase I feasibility project, Primordial Genetics Inc. demonstrated the use of novel template-
independent DNA polymerases (TIDPs) for controlled addition of natural, unmodified nucleotides to
a growing DNA strand in a simple process that does not require a chemical deblocking step. This
innovation has the potential to enable a robust, inexpensive, flexible, environmentally friendly and
easily scalable enzymatic route to manufacturing DNA and RNA used in therapeutics, vaccines,
diagnostics and R&D products.
In this Phase II Small Business Innovation Research (SBIR) project, Primordial Genetics
proposes to continue the Phase I work to optimize the already discovered TIDPs to enable a high
rate (99%) of single nucleotide addition in each synthesis cycle that is needed for an industrial EOS
process. The company’s genetic improvement and screening platform, specifically the Function
Generator™ technology, will be applied together with the artificial intelligence and machine learning
capabilities of our computational biology collaborator Koliber Biosciences to improve these enzymes
to the desired level of efficiency. A prototype EOS process will be developed using the optimized
enzymes acting on oligonucleotides immobilized on a solid support.
The prototype EOS process has the potential to enable more efficient DNA and RNA synthesis
and remove the manufacturing bottlenecks that are currently holding back the development of nucleic
acid medicines. Over time, we will adapt the TIDPs to allow synthesis of all modified nucleotides
currently being incorporated into DNA- and RNA-based therapeutics, vaccines, as well as reagents
for diagnostics, R&D and DNA-based information storage.
