EasyCloneYALI: CRISPR/Cas9‐Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica. Issue 9 (14th February 2018)

• Record Type: Journal Article
• Authors: Holkenbrink, Carina; Dam, Marie I.; Kildegaard, Kanchana R.; Beder, Johannes; Dahlin, Jonathan; Doménech Belda, David; Borodina, Irina
• Is Part Of: Biotechnology journal. Volume 13:Issue 9(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-02-14
• Subjects: CRISPR/Cas9 -- integrative vectors -- metabolic engineering -- Yarrowia lipolytica -- yeast; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201700543
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract : The oleaginous yeast Yarrowia lipolytica is an emerging host for production of fatty acid‐derived chemicals. To enable rapid iterative metabolic engineering of this yeast, there is a need for well‐characterized genetic parts and convenient and reliable methods for their incorporation into yeast. Here, the EasyCloneYALI genetic toolbox, which allows streamlined strain construction with high genome editing efficiencies in Y. lipolytica via the CRISPR/Cas9 technology is presented. The toolbox allows marker‐free integration of gene expression vectors into characterized genome sites as well as marker‐free deletion of genes with the help of CRISPR/Cas9. Genome editing efficiencies above 80% were achieved with transformation protocols using non‐replicating DNA repair fragments (such as DNA oligos). Furthermore, the toolbox includes a set of integrative gene expression vectors with prototrophic markers conferring resistance to hygromycin and nourseothricin. Abstract : EasyCloneYALI is a genome editing toolbox, which allows rapid iterative combinatorial strain engineering in the non‐conventional oleaginous yeast Y. lipolytica . The toolbox allows integration of gene expression cassettes into high expression genome spots with help of selection markers and marker free genome editing using the CRISPR/Cas9 technology.