SBIR-STTR Award

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.

The ability to design and write genomes of living cells provides distinctive opportunities to tackle problems in biomedical, pharmaceutical, agricultural, and chemical industries that are intractable with current technologies. Critically, current state-of-the-art technology is too costly for most researchers to readily access due to the fundamental costs of DNA synthesis chemistry. A new technology is needed to provide inexpensive, rapid, and high-fidelity DNA oligos to the public to meet the challenges rooted in genomic science that can be resolved with synthetic biology. This proposal aims to adapt patented beam pen lithography technology for the construction of an ultra-dense in situ DNA synthesis platform. This patented technology will enable near-field enabled light-directed DNA synthesis, and thereby genomic construction, that is vastly more accessible than current technology, synthesizing at a rate faster than is currently possible while lowering product costs. This will create a paradigm shift that enables large-scale public access to high quality DNA constructs. The proprietary beam pen lithography instrument was adapted to be used in the presence of liquid media and software was developed to integrate automated control over DNA synthesis. Critically, novel microfluidic componentry was developed such that current instrument systems are largely unaffected by the newly integrated microfluidic capabilities. Furthermore, research was conducted to further the ability to conduct photonic PCR as a means of isolating and amplifying tethered oligos of interest using existing BPL and DMD hardware. The continuing optimization of the instrument for DNA synthesis and photonic PCR, with the intent of scaling up output to serve the increasing demand of synthesized DNA to the public as a commercial product is the goal of Phase II. Planned over the course of Phase II is DNA printing and benchmarking against current industry leaders with respect to price, speed, and fidelity, ensuring that the product price would be the most cost effective and reliable on the market. Furthermore, the translation of DNA from the ultra dense oligonucleotide chips into validated and standardized delivery formats is emphasized. Applications for synthesized DNA products span almost every industry, but increased accessibility to this technology will prove crucial to the development of novel research in biotechnology, agriculture and infectious disease prevention. More specifically, it is foreseen that novel synthetic biology products could be used to create resistant forms of plant life for agriculture, develop patient-specific medical treatments based on an individual’s unique genes, and allow rapid screening of new vaccination technologies.