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

The field of spatial biology is poised to provide a wealth of information on the complex intricacies of tissues and symbiotic relationships across the living world. Novel technologies in spatial transcriptomics are producing important insights in human health related to pathology as well as important microscopic relationships that impact our global agriculture, such as plant-fungi interactions. However, for all of the information that is being extracted using these new multi-omics approaches, there is even more that is going unseen as cost limitations and spatial resolution have thus far limited adoption in many research areas. The team aims to continue its work in transforming a patented nanofabrication tool into an unprecedented oligonucleotide synthesis platform capable of generating high density nanoarrays. The work already performed will be the basis of new efforts to commercialize the technology in the field of spatial transcriptomics. This will build on the results of Phase I and Phase II where the team developed a lightdirected DNA synthesis platform capable of subcellular resolution, designed and built a custom high performance digital light engine and developed an automated fluidic delivery system for performing cyclical photochemistries, The work produced new alignment strategies critical to performing high resolution light-based based in liquid environments as well as significant hardware advances in maintaining a leak-proof seal while allowing for micron-scale movements. The work of the project also leads to a new commercial photolithography tool which has since been sold to 5 different research institutions around the world. Continuing these efforts, the Phase IIB project is focused on generating a commercial product that features densely patterned DNA strands featuring unique combinations of barcodes that encode spatial coordinates in the genetic sequence. This product can then be used to extract genetic molecules from a tissue sample and then tag those genetic molecules with those barcodes, effectively placing microscopic longitude and latitude on an organism’s biomolecules. Researchers are then able to create a visual map of what came from where, whether it be DNA, RNA or other related materials. This is an extremely powerful tool for understand how cells respond to each other and to foreign substances, how tumors invade healthy tissue and how cellular organisms interact with other cellular organisms. The applications are far reaching, with potential insights in biofilm fouling in water purification plants, oncology and Alzheimer’s research, chronic illnesses due to toxic exposure and understanding how plants and fungi interact below the soil in our agriculture.