Our ability to efficiently engineer microorganisms to produce desired chemicals at industrially relevant scales, rates, and titers requires a combination of adaptive evolution and rational design methods to screen or select for mutants with optimized phenotypes. While useful, these methods are generally limited by the ability to introduce synthetically designed, non-biased, trackable mutations to clearly identify gene-to-trait relationships. With the advent of CRISPR genetic tools in our favorite model systems such as E. coli or S. cerevisiae, we have made progress towards the development of methods that enabled highly multiplexed and targeted mutagenesis libraries to allow for the rapid identification of genotype-to-phenotype relationships under selective conditions such as growth, production of desired compounds, and tolerance. Unfortunately, the adaptation of these methods for use in non-model systems with desired complex phenotypes that are not easily tractable (e.g. photosynthesis, biomass degradation, CO/CO2 fixation, low pH growth, etc.) is still lacking due to limited basic genetic tools and low transformation/editing efficiencies. In addition, when utilizing CRISPR tools, there can be widely varied success due to a lack of understanding of all of the caveats that affect how different systems will function in each host. I will discuss current progress for the development of expanded CRISPR-based genome engineering tools in some of the non-model microbes we work with in our group to enable accelerated gene-to-trait identification towards improving our understanding of these microbial systems.

Utilizing CRISPR-Cas toolkits for genotype-phenotype discovery and engineering of non-model microbes

ACS Fall 2021

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