ArticleNucleic acids research2021
Recombination machinery engineering facilitates metabolic engineering of the industrial yeast Pichia pastoris.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 79 papers.
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Who cites it
79 citing papers in PubMed.
- Systematic evaluation of the DNA repair machinery facilitated the development of efficient genome editing tools for the red yeastSynthetic and systems biotechnology · 2027Article
- A CRISPR-Cas9 Toolkit Enabling Tunable Integration and Transient Homologous Recombination Enhancement in Yarrowia lipolytica.Biotechnology and bioengineering · 2026Article
- Evolutionary Adaptation and Targeted Metabolic Engineering Synergistically Improve Methanol-Derived Single-Cell Protein Production in Pichia pastoris.Biotechnology journal · 2026Article
- Towards Cost-Effective and Sustainable Media Formulations for Terrestrial and Aquatic Cellular Agriculture.Foods (Basel, Switzerland) · 2026Review
- An advanced genome engineering platform for Komagataella phaffii enabling versatile in vivo DNA assembly and multiplex integration.Journal of biological engineering · 2026Article
- FormatotrophicSynthetic and systems biotechnology · 2026Article
- Expression and identification of a novel high-activity recombinant humanized type I collagen SynthCol1 in Pichia pastoris GS115.Applied microbiology and biotechnology · 2026Article
- rDNA-mediated multicopy integration and gene dosage quantification system for microbial zeaxanthin biosynthesis.World journal of microbiology & biotechnology · 2026Article
- Enhancement of single-stranded template annealing activity by Rad52 during repair of CRISPR-induced dsDNA breaks.Nucleic acids research · 2026Article
- Construction and analysis of a cell factory for terpenoid biosynthesis inSynthetic and systems biotechnology · 2026Article
- CRISPR-based synthetic biology toolkit development inSynthetic and systems biotechnology · 2026Article
- Development of a CRISPR/Cas12a genome editing toolbox inSynthetic and systems biotechnology · 2026Article
- CRISPR/Cas systems in fungal biotechnology: advancing high-value metabolite synthesis for industrial and food security applications.Archives of microbiology · 2026Review
- Benchmarking the PAM compatibility of Cas12a variants for high-throughput yeast genetic variant engineering.Applied and environmental microbiology · 2025Article
- Development of precise genome editing and multi-copy integration tools inSynthetic and systems biotechnology · 2025Article
- Methanol biotransformation for the production of biodegradable plastic monomer L-lactate in yeast.Nature communications · 2025Article
- Efficient production of 2'-fucosyllactose inSynthetic and systems biotechnology · 2025Article
- Promoter engineering for enhanced 3-hydroxypropionic acid production inSynthetic and systems biotechnology · 2025Article
- Design and construction of light-regulated gene transcription and protein translation systems in yeast P. Pastoris.Journal of advanced research · 2025Article
- SELECT: high-precision genome editing strategy via integration of CRISPR-Cas and DNA damage response for cross-species applications.Nucleic acids research · 2025Article
19 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The industrial yeast Pichia pastoris has been harnessed extensively for production of proteins, and it is attracting attention as a chassis cell factory for production of chemicals. However, the lack of synthetic biology tools makes it challenging in rewiring P. pastoris metabolism. We here extensively engineered the recombination machinery by establishing a CRISPR-Cas9 based genome editing platform, which improved the homologous recombination (HR) efficiency by more than 54 times, in particular, enhanced the simultaneously assembly of multiple fragments by 13.5 times. We also found that the key HR-relating gene RAD52 of P. pastoris was largely repressed in compared to that of Saccharomyces cerevisiae. This gene editing system enabled efficient seamless gene disruption, genome integration and multiple gene assembly with positive rates of 68-90%. With this efficient genome editing platform, we characterized 46 potential genome integration sites and 18 promoters at different growth conditions. This library of neutral sites and promoters enabled two-factorial regulation of gene expression and metabolic pathways and resulted in a 30-fold range of fatty alcohol production (12.6-380 mg/l). The expanding genetic toolbox will facilitate extensive rewiring of P. pastoris for chemical production, and also shed light on engineering of other non-conventional yeasts.
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