Rapid Engineering of New Functions in Yeast with Synthetic Chromosomes

Synthetic DNA and modular assembly has revolutionised cellular engineering, allowing new constructs to be built and optimised in days. Through our involvement with the Sc2.0 synthetic yeast project, we are now able to show how a synthetic modular genome can accelerate synthetic biology further. Alongside construction of a 670 kb synthetic yeast chromosome, we have developed protocols to generate diverse pools of yeast strains in hours via the Sc2.0 SCRaMbLE system, which uses Cre/lox recombination to radically change chromosome content and arrangement. In the presence of synthetic constructs, such as those endowing the biosynthesis of pigments or catabolism of xylose, SCRaMbLE can produce 'specialist' host strains overnight with significant improvements in performance in terms of growth and yield. SCRaMbLE can also be used to directly add new genes to Sc2.0 genomes, simply by adding linear synthetic DNA to cells as their chromosomes recombine. With these new modular genome methods together with combinatorial modular DNA construction we have been able to construct a yeast that produces and secretes the beta-lactam antibiotic penicillin.