title: A molecular genetic toolbox forYarrowia lipolytica.
abstract: BACKGROUND
  Yarrowia lipolyticais an ascomycete yeast used in biotechnological research for its abilities to secrete high concentrations of proteins and accumulate lipids. Genetic tools have been made in a variety of backgrounds with varying similarity to a comprehensively sequenced strain.
  
  
  RESULTS
  We have developed a set of genetic and molecular tools in order to expand capabilities ofY. lipolyticafor both biological research and industrial bioengineering applications. In this work, we generated a set of isogenic auxotrophic strains with decreased non-homologous end joining for targeted DNA incorporation. Genome sequencing, assembly, and annotation of this genetic background uncovers previously unidentified genes inY. lipolytica. To complement these strains, we constructed plasmids withY. lipolytica-optimized superfolder GFP for targeted overexpression and fluorescent tagging. We used these tools to build the "Yarrowia lipolyticaCell Atlas," a collection of strains with endogenous fluorescently tagged organelles in the same genetic background, in order to define organelle morphology in live cells.
  
  
  CONCLUSIONS
  These molecular and isogenetic tools are useful for live assessment of organelle-specific protein expression, and for localization of lipid biosynthetic enzymes or other proteins inY. lipolytica. This work provides the Yarrowia community with tools for cell biology and metabolism research inY. lipolyticafor further development of biofuels and natural products.
authors.affiliation: Earth and Biological Sciences Directorate, Environmental Molecular Sciences Laboratory, Richland, WA 99354 USA ; Department of Energy, Battelle EMSL, 3335 Innovation Blvd, Richland, WA 99354 USA.
authors.forename: Erin L
authors.initials: EL
authors.lastname: Bredeweg
authors.affiliation: Chemical & Biological Process Development Group, Energy and Environment Directorate, Pacific Northwest National Laboratories, Richland, WA 99354 USA.
authors.forename: Kyle R
authors.initials: KR
authors.lastname: Pomraning
authors.affiliation: Chemical & Biological Process Development Group, Energy and Environment Directorate, Pacific Northwest National Laboratories, Richland, WA 99354 USA.
authors.forename: Ziyu
authors.initials: Z
authors.lastname: Dai
authors.affiliation: Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden ; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Jens
authors.initials: J
authors.lastname: Nielsen
authors.affiliation: Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden.
authors.forename: Eduard J
authors.initials: EJ
authors.lastname: Kerkhoven
authors.affiliation: Earth and Biological Sciences Directorate, Environmental Molecular Sciences Laboratory, Richland, WA 99354 USA ; Department of Energy, Battelle EMSL, 3335 Innovation Blvd, Richland, WA 99354 USA.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Baker
keywords: GFP localization; Genome sequence; Hygromycin B; Isogenic; Organelle labeling; Overexpression plasmid; Protein tagging; Superfolder GFP; Tools; Yarrowia lipolytica