Toward Rapid and Accessible DNA Synthesis for Synthetic Biology via Beam Pen Lithography

The ability to design and write large DNA fragments provides distinctive opportunities to tackle global problems ranging from food shortages and environmental deterioration to diseases and chemical productions, just to name a few. However, substantial improvements are needed to reduce the cost and increase the speed and reliability of current synthetic genomics tools. TERA-print, LLC has recently developed the world’s first beam pen lithography (BPL) platform, TERA-Fab™ E series, that promises to deliver 10-1000x cheaper synthesis of higher fidelity DNA than with existing technologies in the form of ultra-high-density oligo arrays. This, in turn, has the potential to drive the cost and time of constructing large DNA fragments significantly down not only by making DNA synthesis more accessible but also by shifting the paradigm toward reusable oligo arrays. The ability to pack 10s to 100s of millions of unique oligos per square centimeter opens up the possibility of creating arrays with functional genomes from many organisms that can be amplified and assembled on demand, thereby eliminating the need to resynthesize oligo arrays every time, further reducing the cost and accelerating prototyping of large DNA fragments. In Phase I, TERA-print aims to demonstrate the ability of BPL to produce > 10x higher density and >10x lower cost oligo arrays, as compared to existing technologies. The Company will also quantify the gain in fidelity of oligos synthesized with BPL, owing to the highly localized light intensity at the microprobes that eliminates unintended light exposures and insertion errors, which other photolithographic approaches are prone to. Finally, a rapid and inexpensive method for addressing oligos in the ultra-dense BPL-synthesized arrays using a hybrid approach based on orthogonal primers and photonic polymerase chain reaction (PCR) will be demonstrated that is critical for the development of universal and reusable DNA arrays. These critical demonstrations will put the BPL technology on the map against other DNA synthesis methods and set up the stage for the Phase II development of commercial services, providing the community of synthetic biologists with the ability to order genes and larger fragment DNA with a faster turnaround and at a fraction of the cost than what is available today. New, faster, and cheaper ways to write DNA and engineer biology could have significant impacts on global economies and our personal, daily lives, ranging from health and agriculture to consumer goods, and energy and materials. Gene therapies could offer complete cures of some intractable diseases, such as sickle-cell anemia and epilepsy, crops can be genetically engineered to produce higher yields and be more heat- or drought-resistant, and by rewriting the genetic code, microbes can be tweaked to produce novel materials with unique qualities and new capabilities with the minimum carbon footprint. By bringing new, innovative DNA writing capabilities to synthetic biologists, the proposal aims to fuel this bio-revolution.