(563g) Design Platforms for Modular Cell Engineering and Precise CRISPR Genome Editing of Single and Consortia of Organisms

In this talk, I will present the development of two platform synthetic biology tools in my laboratory. First, to accelerate and reduce the cost of strain engineering, we formulate the modular cell (ModCell) design principles for rapid strain engineering in a plug-and-play fashion. ModCell exploits the modular organization of metabolic networks and combinatorial possibilities of metabolic modules that enable the synthesis of a large space of biochemicals. I will demonstrate ModCell for design, construction, and validation of an Escherichia coli modular cell for combinatorial synthesis of alcohols and bioesters from fermentable sugars and organic wastes. Next, to enable precise CRISPR genome editing of single and consortia of organisms, we have developed the next generation CASPER (CRISPR Associated Software for Pathway Engineering and Research) algorithms and associated software to accommodate the use of CRISPR/Cas systems for genome editing across organisms. CASPER can identify on- and off-targets with enhanced predictability to assist in editing any organism with various endonucleases. Further, CASPER presents novel applications in genome editing: multitargeting analysis (i.e. simultaneous multiple-site modification on a target genome with a gRNA requirement), multispecies population analysis (i.e. gRNA design for genome editing across a microbiome), and PAM prediction. We envision ModCell and CASPER as the useful platform tools for metabolic engineering and synthetic biology applications.