The DOI matches its Crossref record; 11 of the 11 references it cites were found in a registry or an index.
CRISPR/Cas9: a molecular Swiss army knife for simultaneous introduction of multiple genetic modifications in Saccharomyces cerevisiae
Open accessAbstract
A variety of techniques for strain engineering in Saccharomyces cerevisiae have recently been developed. However, especially when multiple genetic manipulations are required, strain construction is still a time-consuming process. This study describes new CRISPR/Cas9-based approaches for easy, fast strain construction in yeast and explores their potential for simultaneous introduction of multiple genetic modifications. An open-source tool (http://yeastriction.tnw.tudelft.nl) is presented for identification of suitable Cas9 target sites in S. cerevisiae strains. A transformation strategy, using in vivo assembly of a guideRNA plasmid and subsequent genetic modification, was successfully implemented with high accuracies. An alternative strategy, using in vitro assembled plasmids containing two gRNAs, was used to simultaneously introduce up to six genetic modifications in a single transformation step with high efficiencies. Where previous studies mainly focused on the use of CRISPR/Cas9 for gene inactivation, we demonstrate the versatility of CRISPR/Cas9-based engineering of yeast by achieving simultaneous integration of a multigene construct combined with gene deletion and the simultaneous introduction of two single-nucleotide mutations at different loci. Sets of standardized plasmids, as well as the web-based Yeastriction target-sequence identifier and primer-design tool, are made available to the yeast research community to facilitate fast, standardized and efficient application of the CRISPR/Cas9 system.
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Cites
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“Therefore, it is logical to speculate that this methodology should have an even higher impact on species of yeasts and filamentous fungi that are notoriously more difficult to alter genetically.”
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Reprogramming Yeast Metabolism from Alcoholic Fermentation to Lipogenesis
Yu, T., Zhou, Y. J., Huang, M., Liu, Q., Pereira, R., David, F., & Nielsen, J. (2018) · Cell · doi:10.1016/j.cell.2018.07.013 · Cited by 341
Potential pitfalls of CRISPR/Cas9‐mediated genome editing
Peng, R., Lin, G., & Li, J. (2016) · The FEBS Journal · doi:10.1111/febs.13586 · Cited by 213
Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae
Generoso, W. C., Gottardi, M., Oreb, M., & Boles, E. (2016) · Journal of Microbiological Methods · doi:10.1016/j.mimet.2016.06.020 · Cited by 165
Acquired multidrug antifungal resistance in Candida lusitaniae during therapy
Asner, S. A., Giulieri, S., Diezi, M., Marchetti, O., & Sanglard, D. (2015) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.02204-15 · Cited by 82
Bayesian genome scale modelling identifies thermal determinants of yeast metabolism
Li, G., Hu, Y., Zrimec, J., Luo, H., Wang, H., Zelezniak, A., Ji, B., & Nielsen, J. (2021) · Nature Communications · doi:10.1038/s41467-020-20338-2 · Cited by 57
Metadata
- Authors
- Mans, R., van Rossum, H. M., Wijsman, M., Backx, A., Kuijpers, N. G. A., van den Broek, M., Daran-Lapujade, P., Pronk, J. T., van Maris, A. J. A., & Daran, J.-M. G.
- Venue
- FEMS Yeast Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2015-03-01
- Volume
- 15
- Issue
- 2
- ISSN
- 1567-1356 (ISSN-L)
- 1567-1364
- OpenAlex
- W2016033442
- Cited by
- 485
- References
- 11
- FWCI
- 19.94
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Mans, R., van Rossum, H. M., Wijsman, M., Backx, A., Kuijpers, N. G. A., van den Broek, M., Daran-Lapujade, P., Pronk, J. T., van Maris, A. J. A., & Daran, J.-M. G. (2015). CRISPR/Cas9: a molecular Swiss army knife for simultaneous introduction of multiple genetic modifications in Saccharomyces cerevisiae. FEMS Yeast Research. https://doi.org/10.1093/femsyr/fov004
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