ArticleScientific reports2017
Development of a comprehensive set of tools for genome engineering in a cold- and thermo-tolerant Kluyveromyces marxianus yeast strain.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 77 citations in OpenAlex.
- Base Editors for Engineering Industrial Microorganisms: Types, Applications, and Future Perspectives.Biotechnology journal · 2026Review
- Development of a CRISPR/Cas12a genome editing toolbox inSynthetic and systems biotechnology · 2026Article
- Metabolic engineering and adaptive laboratory evolution of Kluyveromyces Marxianus for lactic acid production.Microbial cell factories · 2025Article
- Fermentative Yeast Diversity at the Northern Range Limit of Their Oak Tree Hosts.Environmental microbiology reports · 2025Article
- Engineering recombination machinery facilitates the construction of yeast cell factories.FEMS yeast research · 2025Review
- Efficient and markerless gene integration with SlugCas9-HF in Kluyveromyces marxianus.Communications biology · 2024Article
- Comparing the hierarchy of inter- and intra-species interactions with population dynamics of wine yeast cocultures.FEMS yeast research · 2023Article
- RNA polymerase II-driven CRISPR-Cas9 system for efficient non-growth-biased metabolic engineering ofMetabolic engineering communications · 2022Article
- CRISPR-Mediated Base Editing: From Precise Point Mutation to Genome-Wide Engineering in Nonmodel Microbes.Biology · 2022Review
- Considering Strain Variation and Non-Type Strains for Yeast Metabolic Engineering Applications.Life (Basel, Switzerland) · 2022Review
- Protocols for marker-free gene knock-out and knock-down in Kluyveromyces marxianus using CRISPR/Cas9.FEMS yeast research · 2022Article
- BioprospectingFrontiers in bioengineering and biotechnology · 2022Review
- Target-AID-Mediated Multiplex Base Editing in Porcine Fibroblasts.Animals : an open access journal from MDPI · 2021Article
- Improving the functionality of surface-engineered yeast cells by altering the cell wall morphology of the host strain.Applied microbiology and biotechnology · 2021Article
- Growth and autolysis of the kefir yeast Kluyveromyces marxianus in lactate culture.Scientific reports · 2021Article
- Recombination machinery engineering for precise genome editing in methylotrophic yeastiScience · 2021Article
- CRISPR-mediated multigene integration enables Shikimate pathway refactoring for enhanced 2-phenylethanol biosynthesis in Kluyveromyces marxianus.Biotechnology for biofuels · 2021Article
- Advances and Opportunities of CRISPR/Cas Technology in Bioengineering Non-conventional Yeasts.Frontiers in bioengineering and biotechnology · 2021Review
- Genome editing systems across yeast species.Current opinion in biotechnology · 2020Review
- Rational engineering of Kluyveromyces marxianus to create a chassis for the production of aromatic products.Microbial cell factories · 2020Article
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Kluyveromyces marxianus, a non-conventional thermotolerant yeast, is potentially useful for production of ethanol and other products. This species has a strong tendency to randomly integrate transforming DNA fragments, making necessary the development of more precise methods for gene targeting. In this study, we first demonstrated that K. marxianus NBRC1777 is cold-tolerant, and then established a highly efficient and precise technique for gene editing by introducing genes encoding deaminase-mediated targeted point mutagenesis (Target-AID) and clustered regularly interspaced short palindromic repeats (CRISPR) associated proteins (CRISPR-Cas9). We used Target-AID to introduce targeted point mutations that disrupted Nej1 or Dnl4, genes that are involved in non-homologous end-joining (NHEJ). Both of the resulting mutant strains showed enhanced proportions of homology-mediated integration compared to the wild-type parent. In combination with target cleavage by CRISPR-Cas9, markerless integration was performed using short (~50 bp) flanking homologous sequences. Together, these tools render this species fully tractable for gene manipulation, permitting targeted genetic changes in the cold- and thermo-tolerant yeast K. marxianus.
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