EasyCloneYALI : CRISPR/Cas9-Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica
(2018) In Biotechnology Journal 13(9). p.1700543-1700543- 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... (More)
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.
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- author
- Holkenbrink, Carina ; Dam, Marie I LU ; Kildegaard, Kanchana R ; Beder, Johannes ; Dahlin, Jonathan ; Doménech Belda, David and Borodina, Irina
- publishing date
- 2018-09
- type
- Contribution to journal
- publication status
- published
- keywords
- CRISPR-Cas Systems/genetics, Cloning, Molecular/methods, Escherichia coli/genetics, Gene Editing/methods, Metabolic Engineering/methods, Yarrowia/genetics
- in
- Biotechnology Journal
- volume
- 13
- issue
- 9
- pages
- 8 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:29377615
- scopus:85042046691
- ISSN
- 1860-6768
- DOI
- 10.1002/biot.201700543
- language
- English
- LU publication?
- no
- additional info
- © 2018 The Authors. Biotechnology Journal Published by Wiley-VCH Verlag GmbH & Co. KGaA.
- id
- 27a98a47-3787-4218-bc1e-7d5eec47ac2e
- alternative location
- https://onlinelibrary.wiley.com/doi/10.1002/biot.201700543
- date added to LUP
- 2020-12-11 14:46:38
- date last changed
- 2024-06-13 02:47:11
@article{27a98a47-3787-4218-bc1e-7d5eec47ac2e,
abstract = {{<p>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.</p>}},
author = {{Holkenbrink, Carina and Dam, Marie I and Kildegaard, Kanchana R and Beder, Johannes and Dahlin, Jonathan and Doménech Belda, David and Borodina, Irina}},
issn = {{1860-6768}},
keywords = {{CRISPR-Cas Systems/genetics; Cloning, Molecular/methods; Escherichia coli/genetics; Gene Editing/methods; Metabolic Engineering/methods; Yarrowia/genetics}},
language = {{eng}},
number = {{9}},
pages = {{1700543--1700543}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Biotechnology Journal}},
title = {{EasyCloneYALI : CRISPR/Cas9-Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica}},
url = {{http://dx.doi.org/10.1002/biot.201700543}},
doi = {{10.1002/biot.201700543}},
volume = {{13}},
year = {{2018}},
}