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

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.