Construction of efficient cell factories often requires engineering of multi-circuit pathways for synthesis of novel compounds and improved cell productivity. Genome-level screens can identify genes and pathways that enhance the production of desired chemicals, fuels, or biologics. While modern gene editing tools like CRISPR/Cas9 enable precise genome editing, where to integrate new genes and how best to modulate expression remain challenging, especially in non-model organisms. The AltHost Consortium, an MIT-industry partnership, seeks to engineer microbial hosts to be fast, low-cost, and highly productive alternatives for biomanufacturing. Informed by functional genomics, we present tools for modifying non-model organisms for a desired phenotype. First, we demonstrate a framework informed by ATAC-Seq and RNA-Seq for identifying optimal landing sites for heterologous expression cassettes. Second, we present a tool based on the Kozak sequence for tuning protein expression to a target level without promoter engineering. We demonstrate the utility of these tools by engineering the yeast <i>P. pastoris</i> to be a highly productive cell factory for quality biologics. By leveraging functional genomics, we believe our approach can be extended to develop these tools in almost any microbial host.

Universal tools informed by functional genomics for constructing microbial cell factories

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.