ArticleScientific reports2018
Genome-wide prediction of CRISPR/Cas9 targets in Kluyveromyces marxianus and its application to obtain a stable haploid strain.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Yeast species resource exploration and application in cell factories.Synthetic and systems biotechnology · 2026Review
- Development of a CRISPR/Cas12a genome editing toolbox inSynthetic and systems biotechnology · 2026Article
- Expression of EndonucleaseCurrent issues in molecular biology · 2026Article
- Creating a large designer cellulosome in yeast to boost ethanol production.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Metabolic engineering and adaptive laboratory evolution of Kluyveromyces Marxianus for lactic acid production.Microbial cell factories · 2025Article
- Gene disruption via a transient hypercompact CRISPR-AsCas12f1 system in Kluyveromyces marxianus.Biotechnology letters · 2025Article
- From genetic constraints to process hurdles: A review ofCurrent research in food science · 2025Review
- RNA polymerase II-driven CRISPR-Cas9 system for efficient non-growth-biased metabolic engineering ofMetabolic engineering communications · 2022Article
- Protocols for marker-free gene knock-out and knock-down in Kluyveromyces marxianus using CRISPR/Cas9.FEMS yeast research · 2022Article
- Insights on life cycle and cell identity regulatory circuits for unlocking genetic improvement in Zygosaccharomyces and Kluyveromyces yeasts.FEMS yeast research · 2021Article
- Meiosis-Based Laboratory Evolution of the Thermal Tolerance inFrontiers in bioengineering and biotechnology · 2021Article
- Genome editing systems across yeast species.Current opinion in biotechnology · 2020Review
- Gross Chromosomal Rearrangements inInternational journal of molecular sciences · 2020Article
- Valorisation of pectin-rich agro-industrial residues by yeasts: potential and challenges.Applied microbiology and biotechnology · 2020Review
- Constructing a yeast to express the largest cellulosome complex on the cell surface.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Article
- Constructing a human complex type N-linked glycosylation pathway in Kluyveromyces marxianus.PloS one · 2020Article
- Biological Parts forFrontiers in bioengineering and biotechnology · 2019Article
- Article
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8 authors.
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Abstract
Kluyveromyces marxianus, a probiotic yeast, is important in industrial applications because it has a broad substrate spectrum, a rapid growth rate and high thermotolerance. To date, however, there has been little effort in its genetic engineering by the CRISPR/Cas9 system. Therefore, we aimed at establishing the CRISPR/Cas9 system in K. marxianus and creating stable haploid strains, which will make genome engineering simpler. First, we predicted the genome-wide target sites of CRISPR/Cas9 that have been conserved among the eight sequenced genomes of K. marxianus strains. Second, we established the CRISPR/Cas9 system in the K. marxianus 4G5 strain, which was selected for its high thermotolerance, rapid growth, a pH range of pH3-9, utilization of xylose, cellobiose and glycerol, and toxin tolerance, and we knocked out its MATα3 to prevent mating-type switching. Finally, we used K. marxianus MATα3 knockout diploid strains to obtain stable haploid strains with a growth rate comparable to that of the diploid 4G5 strain. In summary, we present the workflow from identifying conserved CRISPR/Cas9 targets in the genome to knock out the MATα3 genes in K. marxianus to obtain a stable haploid strain, which can facilitate genome engineering applications.
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