The colors of CRISPR-Cas Type V: Broadening the genome editing toolkit for Saccharomyces cerevisiae

Samenvatting (Engelstalig)

The use of microorganisms for biotechnological processes allows the natural and heterologous production of industrially relevant compounds. Genome editing tools are required to incorporate heterologous biosynthetic production pathways or to fine-tune already existing metabolic pathways. The application of CRISPR-Cas systems in prokaryotes and eukaryotes has revolutionized genome editing programs in the last years. Natural diversity of these CRISPR-Cas systems allows the development of novel tools with different characteristics, which can grant higher editing efficiencies, integration of large DNA constructs or access to novel targets. Moreover, modification of characterized tools can facilitate genome editing of difficult-to-modify organisms, including industrially relevant strains. In this thesis, we explored the diversity of type V CRISPR-Cas effectors and performed some first steps towards the application of several nucleases in the industrially relevant yeast Saccharomyces cerevisiae. We evaluated their activity and proposed modifications to already existing prokaryotic tools for their use in eukaryotic systems.