Highlights of our work include developing a novel systematic mutagenesis method (Higgins et al. Much of this effort has been focused on leveraging recent advances in DNA sequencing to interrogate protein function in higher throughput and also demonstrating that protein sequence topology is a critical, yet often unexplored, dimension of the sequence-function landscape (Higgins and Savage 2017). We therefore also develop genetic tools to interrogate proteins in the cell. That is, we are interested not only in protein mechanism but also in trying to understand this mechanism in its native, cellular context. Our group takes an integrative view of protein function. Tools for Investigating Protein Structure, Function and Engineering The latter publication is the first such successful demonstration of a fully functional, engineered CCM. 2019), and in a recent paper, demonstrated a complete reconstitution of the CCM by porting it from a native to a heterologous host (Flamholz et al. We have used high-throughput screening strategies to identify unknown components of the CCM (Desmarais et al. We are therefore also developing tractable genetic systems to explore, in a synthetic biological context, how CCMs can be constructed de novo. Second, work from our group has demonstrated the inherent genetic modularity of the CCM may all for it to be transferred to other organisms in order to improve their growth (Bonacci et al. 2016) suggests that pH, carbon transport, and carboxysome permeability are highly intertwined and we are interested in mechanisms that can control the permeability of the carboxysome shell. For example, our systems model of the CCM (Mangan et al. Despite numerous structural studies of carboxysome components, there are many open questions we are currently investigating related to how this massive complex self-assembles and functions in the context of the cellular milieu. Rubisco is a notoriously bad enzyme, and it is thought the inside of the carboxysome provides a microenvironment within the cell of high carbon dioxide (and possibly low oxygen) as a means of improving rate and specificity. Perhaps most amazing is the use of a protein organelle, the carboxysome, to achieve improved activity of Rubisco. Understanding and Improving Biological CO 2 AssimlationĬyanobacteria carry out photosynthetic carbon dioxide assimilation by controlling when and where these biochemical reactions occur. Our specific areas of research are as follows. Given the generality of these tools, we also explore how engineered genome editors can be used to improve editing in human cells. We are therefore also interested in the development of novel genome editing technologies for accelerating genetic experiments in plants.
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