Progress in DNA sequencing has occurred through multiple stages of disruptive new technologies being introduced to the field, each of which has increased sequencing capabilities by lowering costs, improving throughput, and reducing errors. The goal of this research project is to investigate a new, all-electronic sequencing method that has the potential to become the next transformative step for DNA sequencing. This new method is based on single DNA polymerase molecules bound to nanoscale electronic transistors, a hybrid device that transduces the activity of a single polymerase molecule into an electronic signal. The goal of this research project is to determine whether these hybrid polymerase-transistors are truly applicable to DNA sequencing and the competitive environment of advanced sequencing technologies. To answer this question, the project teams the scientists who have developed the devices with Illumina, Inc., a worldwide leader in the DNA sequencing market. The experiments proposed here build on encouraging preliminary results, first to demonstrate accurate DNA sequencing and second to evaluate whether the new technique could become a competitive challenge to other sequencing methods. The interdisciplinary team will combine state-of-the-art techniques from protein engineering, nanoscale fabrication, and machine learning to customize polymerase's activity and its interactions with the electronic transistors. If successful, nanoscale solid-state devices like transistors provide one of the best opportunities for increasing sequencing capabilities while decreasing sequencing costs, so that DNA sequencing can become a standard technique in health care and disease treatment.
Over the past two decades, DNA sequencing has transformed from a heroic, nearly impossible task to a routine component of modern laboratory research. The field of DNA sequencing has improved tremendously through a strategy of modifying and monitoring polymerases, a key enzyme at the heart of many DNA sequencing technologies. This proposal is motivated by developments in the field of single-molecule electronics, which provide an entirely new mode for listening to the activity of single polymerase molecules. This electronic method is very different from the biochemical, optical, or nanopore-based techniques currently in use, and it has inherent advantages that could provide exciting possibilities for DNA sequencing. The project will tailor single-molecule electronics for the specific purpose of DNA sequencing and determine whether this strategy could lead to a new generation of sequencing technology.
| Richardson, Mark B; Brown, Derek B; Vasquez, Carlos A et al. (2018) Synthesis and Explosion Hazards of 4-Azido-l-phenylalanine. J Org Chem 83:4525-4536 |
| Pugliese, Kaitlin M; Weiss, Gregory A (2017) Recent progress in dissecting molecular recognition by DNA polymerases with non-native substrates. Curr Opin Chem Biol 41:43-49 |
