ArticleBMC microbiology2014
Molecular characterization of KU70 and KU80 homologues and exploitation of a KU70-deficient mutant for improving gene deletion frequency in Rhodosporidium toruloides.
Article in BMC microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 45 papers.
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45 citing papers in PubMed, 122 citations in OpenAlex.
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- Engineering Rhodotorula toruloides as a platform organism for de novo synthesis of fatty-acid esters.Nature communications · 2026Article
- Establishment of a protoplast transformation method for the oleaginous yeast Rhodotorula toruloides.FEMS yeast research · 2026Article
- The role of ATP citrate lyase, phosphoketolase, and malic enzyme in oleaginous Rhodotorula toruloides.Applied microbiology and biotechnology · 2025Article
- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Engineering of xylose metabolic pathways in Rhodotorula toruloides for sustainable biomanufacturing.FEMS yeast research · 2025Review
- Engineering recombination machinery facilitates the construction of yeast cell factories.FEMS yeast research · 2025Review
- Current advances in alteration of fatty acid profile in Rhodotorula toruloides: a mini-review.World journal of microbiology & biotechnology · 2023Review
- Establishment of a CRISPR/Cas9-Mediated Efficient Knockout System ofJournal of fungi (Basel, Switzerland) · 2023Article
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- Establishment of High-Efficiency Screening System for Gene Deletion inJournal of fungi (Basel, Switzerland) · 2022Article
- Knockout of a highly GC-rich gene in Burkholderia pyrrocinia by recombineering with freeze-thawing transformation.Molecular plant pathology · 2021Article
- Engineering oleaginous yeast Rhodotorula toruloides for overproduction of fatty acid ethyl esters.Biotechnology for biofuels · 2021Article
- Understanding and exploiting the fatty acid desaturation system in Rhodotorula toruloides.Biotechnology for biofuels · 2021Article
- Biodiesel Production From Lignocellulosic Biomass Using Oleaginous Microbes: Prospects for Integrated Biofuel Production.Frontiers in microbiology · 2021Review
- Editorial: Physiology, Application, and Bioengineering of Oleaginous Microorganisms.Frontiers in microbiology · 2021Article
- The High Osmolarity Glycerol (HOG) Pathway Functions in Osmosensing, Trap Morphogenesis and Conidiation of the Nematode-Trapping FungusJournal of fungi (Basel, Switzerland) · 2020Article
- Rhodosporidium toruloides - A potential red yeast chassis for lipids and beyond.FEMS yeast research · 2020Review
- Consolidated bioprocessing for butanol production of cellulolytic Clostridia: development and optimization.Microbial biotechnology · 2020Review
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundRhodosporidium toruloides is a β-carotenoid accumulating, oleaginous yeast that has great biotechnological potential. The lack of reliable and efficient genetic manipulation tools have been a major hurdle blocking its adoption as a biotechnology platform.
resultsWe report for the first time the development of a highly efficient targeted gene deletion method in R. toruloides ATCC 10657 via Agrobacterium tumefaciens-mediated transformation. To further improve targeting frequency, the KU70 and KU80 homologs in R. toruloides were isolated and characterized in detail. A KU70-deficient mutant (∆ku70e) generated with the hygromycin selection cassette removed by the Cre-loxP recombination system showed a dramatically improved targeted gene deletion frequency, with over 90% of the transformants being true knockouts when homology sequence length of at least 1 kb was used. Successful gene targeting could be made with homologous flanking sequences as short as 100 bp in the ∆ku70e strain. KU70 deficiency did not perturb cell growth although an elevated sensitivity to DNA mutagenic agents was observed. Compared to the other well-known oleaginous yeast, Yarrowia lipolytica, R. toruloides KU70/KU80 genes contain much higher density of introns and are the most GC-rich KU70/KU80 genes reported.
conclusionsThe KU70-deficient mutant generated herein was effective in improving gene deletion frequency and allowed shorter homology sequences to be used for gene targeting. It retained the key oleaginous and fast growing features of R. toruloides. The strain should facilitate both fundamental and applied studies in this important yeast, with the approaches taken here likely to be applicable in other species in subphylum Pucciniomycotina.
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