ArticleFEMS yeast research2018
Genome editing in Kluyveromyces and Ogataea yeasts using a broad-host-range Cas9/gRNA co-expression plasmid.
Article in FEMS yeast research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 49 papers.
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Who cites it
49 citing papers in PubMed, 158 citations in OpenAlex.
- Microbial Hydrolysates as Amino Acid Source in Cell Culture Media for Cellular Agriculture.Biotechnology and bioengineering · 2026Review
- Mitochondrial localization of key NADApplied and environmental microbiology · 2026Article
- Development of a CRISPR/Cas12a genome editing toolbox inSynthetic and systems biotechnology · 2026Article
- Development of precise genome editing and multi-copy integration tools inSynthetic and systems biotechnology · 2025Article
- Yeast Midas' touch: recent advances in the valorisation of methanol by yeasts.World journal of microbiology & biotechnology · 2025Review
- Uncoupling protein production from growth: different strategies for intracellular and secreted proteins in yeast.Microbial cell factories · 2025Article
- Exploring Yeast's Energy Dynamics: The General Stress Response Lowers Maintenance Energy Requirement.Microbial biotechnology · 2025Article
- Functional analysis of Saccharomyces cerevisiae FLO genes through optogenetic control.FEMS yeast research · 2025Article
- Lignocellulosic biomass fermentation: a roadmap for Candida famata and Ogataea polymorpha.FEMS yeast research · 2025Review
- Multi-gene precision editing tool using CRISPR-Cas12a/Cpf1 system in Ogataea polymorpha.Microbial cell factories · 2025Article
- Screening of broad-host expression promoters for shuttle expression vectors in non-conventional yeasts and bacteria.Microbial cell factories · 2024Article
- Efficient and markerless gene integration with SlugCas9-HF in Kluyveromyces marxianus.Communications biology · 2024Article
- Multiplex CRISPR-Cas Genome Editing: Next-Generation Microbial Strain Engineering.Journal of agricultural and food chemistry · 2024Review
- Physiological relevance, localization and substrate specificity of the alternative (type II) mitochondrial NADH dehydrogenases ofFrontiers in microbiology · 2024Article
- The biological principles and advanced applications of DSB repair in CRISPR-mediated yeast genome editing.Synthetic and systems biotechnology · 2023Review
- Expanding the genome editing toolbox of Saccharomyces cerevisiae with the endonuclease ErCas12a.FEMS yeast research · 2023Article
- CRISPR-Cas9 engineering in the hybrid yeast Zygosaccharomyces parabailii can lead to loss of heterozygosity in target chromosomes.FEMS yeast research · 2023Article
- Metabolic engineering of non-conventional yeasts for construction of the advanced producers of biofuels and high-value chemicals.BBA advances · 2023Article
- RNA polymerase II-driven CRISPR-Cas9 system for efficient non-growth-biased metabolic engineering ofMetabolic engineering communications · 2022Article
- Identification of a novel gene required for competitive growth at high temperature in the thermotolerant yeastMicrobiology (Reading, England) · 2022Article
Corrections and comments
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Authors and funding
11 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
While CRISPR-Cas9-mediated genome editing has transformed yeast research, current plasmids and cassettes for Cas9 and guide-RNA expression are species specific. CRISPR tools that function in multiple yeast species could contribute to the intensifying research on non-conventional yeasts. A plasmid carrying a pangenomic origin of replication and two constitutive expression cassettes for Cas9 and ribozyme-flanked gRNAs was constructed. Its functionality was tested by analyzing inactivation of the ADE2 gene in four yeast species. In two Kluyveromyces species, near-perfect targeting (≥96%) and homologous repair (HR) were observed in at least 24% of transformants. In two Ogataea species, Ade- mutants were not observed directly after transformation, but prolonged incubation of transformed cells resulted in targeting efficiencies of 9% to 63% mediated by non-homologous end joining (NHEJ). In an Ogataea parapolymorpha ku80 mutant, deletion of OpADE2 mediated by HR was achieved, albeit at low efficiencies (<1%). Furthermore the expression of a dual polycistronic gRNA array enabled simultaneous interruption of OpADE2 and OpYNR1 demonstrating flexibility of ribozyme-flanked gRNA design for multiplexing. While prevalence of NHEJ prevented HR-mediated editing in Ogataea, such targeted editing was possible in Kluyveromyces. This broad-host-range CRISPR/gRNA system may contribute to exploration of Cas9-mediated genome editing in other Saccharomycotina yeasts.
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