Enabling a robust biodesign tool box for the oleaginous yeast, Yarrowia lipolytica

Yarrowia lipolytica is a genetically tractable, genome sequenced oleaginous yeast. In order to accelerate the development of strains for biofuel and bioproduct production, we have generated a molecular genetic "tool box" for this organism. The foundation of this tool box are single or double auxotrophic strains with a deletion of the non-homologous end joining (NHEJ) DNA repair pathway. Elimination of NHEJ facilitates targeted gene tagging, overexpression and disruption. In this genetic background we have developed a cell atlas, with GFP tagged proteins that target the nucleus, mitochondria, lipid droplet, endoplasmic reticulum, golgi, vacuole and peroxisome. Using the cell atlas we are able to monitor organelles under nitrogen limiting lipid production conditions in gene overexpression, deletion and wildtype strains. Finally, we are utilizing global metabolomic and proteomic analyses in order to identify pathways and regulatory genes for genetic manipulation. With our strains, we monitor the biological processes Y. lipolytica in time and space.