(4cf) Synthetic Biology Tools for Protein and Genome Engineering

Rapidly advancing technologies like multiplex oligonucleotide synthesis and next-generation high-throughput sequencing have opened the door to tremendous progress in synthetic biology. My experience in both protein engineering and strain engineering puts me in an excellent position to forge the next generation of synthetic biology tools for protein and genome engineering.

My research career started as an undergraduate at Johns Hopkins University under Prof. Marc Ostermeier. I helped engineer allosteric molecular switches via directed evolution (Guntas, Mansell, et al., PNAS, 2005).  During my PhD (with Prof. Matthew DeLisa, Cornell), I developed a suite of tools for examining protein folding, protein-protein interactions, and post-translational modifications in E. coli. I developed a protein folding reporter for the bacterial periplasm (Mansell et al., Protein Science, 2010) using a Beta-lactamase (BLA) based selection. I showed that we could not only probe intrinsic protein folding, but factors in the cellular environment (e.g., chaperones, redox state) that affected protein folding.  In addition, I optimized a system for detecting protein-protein interactions in the periplasm using split BLA.  Finally, I refactored the protein folding reporter to function as a detector for glycosylation in bacteria (Mansell et al., Biotech. J., 2013) as a pipeline for engineering post-translational modifications. Basically, I was in the business of selection design.

My postdoctoral work with Prof. Ryan Gill at the University of Colorado Boulder gave me a broader view of both metabolic engineering and the design of directed evolution experiments.  By combining multiplex oligonucleotide synthesis and recombineering, I developed a platform for directed evolution of orthogonal repressors for cellular circuits on both the DNA and RNA level. My postdoctoral work has given me a unique perspective on generating targeted libraries, generating programmed diversity to complement my expertise in selection design.

My future research directions will combine protein engineering and strain engineering to produce high-value products in bacteria. I am particularly interested in the effect of post-translational modifications on protein stability and function.  Multiplex recombineering and high-throughput sequencing allow us to glean an unprecedented amount of information about structure-activity relationships, including the engineered placement of post-translational modifications in a protein. Multiplexed synthesis allows for heretofore unapproachable pathway engineering opportunities as well.  By combining these molecular toolkits, we can develop platforms to engineer both native hosts and exogenous pathways for the production of therapeutic proteins, biofuels, and other high-value products using programmed diversity and smart selections.

Finally, I have a particular interest in teaching, especially in teaching biology to chemical engineering students. I have been trained in applying active learning techniques in the classroom to keep students engaged and involved in the material. Active and cooperative learning will be central to whichever courses I teach.  As Niels Bohr once supposedly said: "Biology is too important to be left to the biologists."