Yeast 2.0 and beyond: building the world’s first synthetic eukaryote

Paulsen IT and The Australian Team For Yeast 2.0

Macquarie University, Sydney Australia.

Yeast 2.0 is an international consortium aiming to build the world’s first synthetic eukaryote by 2017. Systematic genome wide changes in the synthetic yeast include TAG/TAA stop-codon replacements, deletion of subtelomeric regions, introns, transfer RNAs, transposons, and insertion of loxPsym recombination sites. The Australian Yeast 2.0 team is responsible for the design and synthesis of synthetic versions of chromosomes 14 and 16. Construction is essentially complete at Macquarie, with 100% of the synthetic DNA successfully inserted. Troubleshooting and repairing errors identified through genome sequencing is currently ongoing. One of the most interesting features of Yeast 2.0 is the incorporation of the SCRaMbLE system for generating combinatorial genomic diversity through rearrangements at loxPsym recombination sites. This opens up the possibility of harnessing the SCRaMbLE system for adaptive laboratory evolution experiments. We have developed biosensors that respond to a variety of industrially useful metabolites, and are now seeking to use a combination of SCRaMbLE-ing and flow cytometry to identify strains that can produce higher levels of these metabolites.