Exploring the potential of yeast mitochondria for synthetic cell research

Building synthetic cells is extremely interesting from a fundamental perspective, as the ability to rationally build viable, dividing, and self-maintaining cells provides knowledge on the minimal requirements to sustain life. This should enhance our understanding about which biological parts are minimally required for a cell to live, and how different cellular functions work and interact with each other. Besides a fundamental understanding of life, synthetic cells can also be applied as synthetic biology tools, for example for synthesis and delivery of therapeutics, or the production of compounds that cannot be produced with currently available organisms used as cell factories. The overarching goal of the research described in this thesis was to devise a strategy for building genomes for synthetic cell, using baker’s yeast Saccharomyces cerevisiae. Two methods were explored, the first being building a genome de novo using yeast in vivo assembly, and secondly, it was investigated whether the preexisting minimal genome of S. cerevisiae mitochondria could be expanded. To this end, yeast mitochondrial DNA and RNA was characterized using novel methods and various strategies for engineering the yeast mitochondrial genome were tested.