(6cq) Expanding the Chemistry of Life for the Development of Novel Therapeutics and Synthetic Organisms

Synthetic biology has a fundamental scientific focus on developing biological parts to understand and manipulate biological systems. My multidisciplinary research program will focus on utilizing expansions in the chemistry of genetically encoded systems at both the protein and nucleic acids level for the development of engineered biological systems.  Aim1: At the protein level, my work will focus on the co-translational incorporation of L-phosphoserine (Sep) into proteins. Our biochemical understanding of protein phosphorylation has been fundamentally limited by our technological inability to manufacture useful quantities of homogenous, site-specific phosphoserylated human proteins. The platform I have developed during my postdoctoral research now allows robust production of active human phosphoproteins. I will use this capacity to address previously intractable questions in cellular signaling associated with epigenetic regulation and disease progression. This work has the potential to add breadth and depth to our understanding as well as provide novel therapeutic windows.  Aim 2: While engineered biological components display structural and functional orthogonally to their natural counterparts, they lack chemical orthogonality. To overcome this challenge, I will focus on expanding nucleic acid biochemistry by integrating 2 additional letters to the genetic code. My work will utilize synthetic nucleobases imidazoaminopyrimidone (letter: P) and nitroaminopyridone (letter: Z) in conjunction with G, A, T, and C to develop synthetic DNA and RNA ‘parts’ for biological engineering applications. P and Z form Watson-Crick pairing like G-C and A-T, do not significantly alter DNA stereochemistry, and are compatible with fundamental biochemical processes such as DNA replication and transcription. The methods I developed during my postdoctoral research allow for assembling P/Z containing synthetic genes. I will use this chemical expansion of the genetic code to develop modular synthetic gene components such as promoters, terminators, ribosome binding sites, riboswitches, as well as chemically-orthogonal translation systems for non-standard amino acid incorporation. In the long run, such biological ‘parts’ displaying chemical orthogonality provided by the P:Z pair could play an integral role in the bio-containment of synthetic organisms. These efforts will be sustained by my training in RNA-protein biochemistry and augmented by my experience in biological engineering. Furthermore, these aims will provide a framework for training Masters’ and PhD level students, and will complement my interest in teaching undergraduate students.