METAL - Enzymatic Photolithography

The chemical synthesis of DNA, RNA and other types of nucleic acids is a highly efficient and cost- effective process that has been adopted by the overwhelming majority of academic and industrial players. This process, employing highly reactive DNA and RNA building blocks called phosphoramidites and with the DNA chain growing one building block at a time on small beads, remains virtually unchanged more than fourty years later since its inception. A notable development has been the ability to synthesize DNA molecules in parallel with minor modifications of phosphoramidite chemistry, producing objects called microarrays. Microarrays, or high-throughput nucleic acid synthesis, can meet the ever-growing demand for longer, more complex sequence libraries in research venues such as data storage on DNA, gene synthesis, and synthetic biology. Meanwhile, recently, a new strategy emerged for the synthesis of nucleic acids, one that employs polymerases to grow nucleic acids chains. Important, those polymerases do not require a so-called template, ie a DNA strand, to produce the complement of, like those involved in DNA replication and RNA transcription. Rather, these enzymes extend DNA chains in an untemplated manner, randomly adding nucleotides. In order to grow chains of the desired sequence, most enzymatic approaches have come up with a chemical solution to add nucleotides in a stepwise manner, blocking further enzymatic extension until an appropriate reagent unblocks the chain for the addition of the next nucleotide. Enzymatic nucleic acid synthesis holds the promise to bypass the limitations of phosphoramidite chemistry, achieving greater chain length at lower production costs and with a greatly reduced environment footprint. In this project, we want to combine these two recent developments enzymatic synthesis and microarray fabrication into a single manufacturing platform where DNA and RNA synthesis takes place enzymatically and in a parallel manner so as to produce hundreds of thousands of sequences in a single run. This project is an Austria-France international collaboration between the groups of Jory Lietard at the University of Vienna and Fabienne Levi-Acobas at Pasteur Institute where they will join their respective forces and expertise in the field of microarray and enzymatic synthesis to make that project a reality.