ArticleApplied and environmental microbiology2014
Construction of a quadruple auxotrophic mutant of an industrial polyploid saccharomyces cerevisiae strain by using RNA-guided Cas9 nuclease.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 65 papers.
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65 citing papers in PubMed.
- Systematic design of auxotrophic strains and media conditions to probe metabolic functions in E. coli.PLoS computational biology · 2026Article
- Response mechanisms of xylitol-producing Saccharomyces cerevisiae strains to acetic acid and furfural and the role of SPI1, CTT1, and CLB1 in tolerance.Microbial cell factories · 2026Article
- Improving inhibitor tolerance of xylitol-producing Saccharomyces cerevisiae by overexpressing key target genes mined through comparative transcriptomes.Biotechnology for biofuels and bioproducts · 2026Article
- Alleviation of ER stress via targeted genetic engineering enhances recombinant ovalbumin secretion inFood science and biotechnology · 2026Article
- Scarless Genetic Engineering ofJournal of microbiology and biotechnology · 2025Article
- Enhanced heme production in industrial Saccharomyces cerevisiae through metabolic engineering.NPJ science of food · 2025Article
- Engineering Saccharomyces boulardii for enhanced surface display capacity.Microbial cell factories · 2025Article
- Synthetic evolution ofmLife · 2025Review
- Assessing the Presence of Phosphoinositides on Autophagosomal Membrane in Yeast by Live Cell Imaging.Microorganisms · 2024Article
- Conversion of polyploid and alloploid Saccharomyces sensu stricto strains to leu2 mutants by genome DNA editing.Applied microbiology and biotechnology · 2024Article
- Construction of an alternative NADPH regeneration pathway improves ethanol production inSynthetic and systems biotechnology · 2024Article
- Towards a hybrid model-driven platform based on flux balance analysis and a machine learning pipeline for biosystem design.Synthetic and systems biotechnology · 2024Article
- Novel Roles of the Greatwall Kinase Rim15 in Yeast Oxidative Stress Tolerance through Mediating Antioxidant Systems and Transcriptional Regulation.Antioxidants (Basel, Switzerland) · 2024Article
- Development of a Novel ATP Bioluminescence Assay Based on Engineered ProbioticJournal of microbiology and biotechnology · 2023Article
- CRISPR-Cas Technology for Bioengineering Conventional and Non-Conventional Yeasts: Progress and New Challenges.International journal of molecular sciences · 2023Review
- Switching carbon metabolic flux for enhancing the production of sesquiterpene-based high-density biofuel precursor in Saccharomyces cerevisiae.Biotechnology for biofuels and bioproducts · 2023Article
- Practical Approaches for the YeastInternational journal of molecular sciences · 2023Review
- Deletion of QDR genes in a bioethanol-producing yeast strain reduces propagation of contaminating lactic acid bacteria.Scientific reports · 2023Article
- Alleviating glucose repression and enhancing respiratory capacity to increase itaconic acid production.Synthetic and systems biotechnology · 2023Article
- ACtivE: Assembly and CRISPR-TargetedACS synthetic biology · 2022Article
5 more citing papers are in PubMed but not listed here.
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6 authors.
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Abstract
Industrial polyploid yeast strains harbor numerous beneficial traits but suffer from a lack of available auxotrophic markers for genetic manipulation. Here we demonstrated a quick and efficient strategy to generate auxotrophic markers in industrial polyploid yeast strains with the RNA-guided Cas9 nuclease. We successfully constructed a quadruple auxotrophic mutant of a popular industrial polyploid yeast strain, Saccharomyces cerevisiae ATCC 4124, with ura3, trp1, leu2, and his3 auxotrophies through RNA-guided Cas9 nuclease. Even though multiple alleles of auxotrophic marker genes had to be disrupted simultaneously, we observed knockouts in up to 60% of the positive colonies after targeted gene disruption. In addition, growth-based spotting assays and fermentation experiments showed that the auxotrophic mutants inherited the beneficial traits of the parental strain, such as tolerance of major fermentation inhibitors and high temperature. Moreover, the auxotrophic mutants could be transformed with plasmids containing selection marker genes. These results indicate that precise gene disruptions based on the RNA-guided Cas9 nuclease now enable metabolic engineering of polyploid S. cerevisiae strains that have been widely used in the wine, beer, and fermentation industries.
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