Acceleration of cell factories engineering using CRISPR-based technologies

The constant demand of oil-derived products in the market has pushed science to develop alternative ways to cope with this demand. Therefore the development of efficient cell factories as sustainable alternative is an expanding trend. These are envisioned as future workhorse manufacturers of pharmaceuticals, biofuels and biomaterials. The focus of this thesis is to develop new genome engineering methods to relieve one of the major bottlenecks in metabolic engineering, the strain design and optimization. The aim is to generate an engineering tool-box applicable to different model organisms, which can potentially be standardized in an automatable platform and, in the future be integrated with metabolic modeling tools. In particularly it describes the technologies developed in the three widely used
organisms: E. coli, S. cerevisiae and CHO mammalian cells using the recent breakthrough CRISPR/ Cas9 system. These include CRMAGE, a MAGE improved recombineering platform using CRISPR negative selection, CrEdit, a system for multi-loci marker-free simultaneous gene and pathway integrations and CRISPy a platform to accelerate genome editing in CHO cells.