title: Synthetic biology and molecular genetics in non-conventional yeasts: Current tools and future advances.
abstract: Coupling the tools of synthetic biology with traditional molecular genetic techniques can enable the rapid prototyping and optimization of yeast strains. While the era of yeast synthetic biology began in the well-characterized model organism Saccharomyces cerevisiae, it is swiftly expanding to include non-conventional yeast production systems such as Hansenula polymorpha, Kluyveromyces lactis, Pichia pastoris, and Yarrowia lipolytica. These yeasts already have roles in the manufacture of vaccines, therapeutic proteins, food additives, and biorenewable chemicals, but recent synthetic biology advances have the potential to greatly expand and diversify their impact on biotechnology. In this review, we summarize the development of synthetic biological tools (including promoters and terminators) and enabling molecular genetics approaches that have been applied in these four promising alternative biomanufacturing platforms. An emphasis is placed on synthetic parts and genome editing tools. Finally, we discuss examples of synthetic tools developed in other organisms that can be adapted or optimized for these hosts in the near future.
authors.affiliation: McKetta Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St., Stop C0400, Austin, TX 78712, United States.
authors.forename: James M
authors.initials: JM
authors.lastname: Wagner
authors.affiliation: McKetta Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St., Stop C0400, Austin, TX 78712, United States; Institute for Cellular and Molecular Biology, The University of Texas at Austin, 2500 Speedway Avenue, Austin, TX 78712, United States. Electronic address: halper@che.utexas.edu.
authors.forename: Hal S
authors.initials: HS
authors.lastname: Alper
mesh_terms: D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D000072669:Gene Editing; D008967:Molecular Biology / Q000379:methods*; D010843:Pichia / Q000235:genetics; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D058615:Synthetic Biology / Q000379:methods*; D025062:Yarrowia / Q000235:genetics; D015003:Yeasts / Q000235:genetics*
keywords: CRISPR; Cas9; Hansenula polymorpha; Kluyveromyces lactis; Pichia pastoris; Synthetic biology; Yarrowia lipolytica; Yeast