ArticleBMC systems biology2017
GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris.
Article in BMC systems biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 88 papers.
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Who cites it
88 citing papers in PubMed, 189 citations in OpenAlex.
- Article
- Functional In Vitro Expression of the Succinate-Acetate Transporter SatP and an Initial Screening Strategy.Membranes · 2026Article
- An advanced genome engineering platform for Komagataella phaffii enabling versatile in vivo DNA assembly and multiplex integration.Journal of biological engineering · 2026Article
- Evaluation of recombinant trypsinogen and chymotrypsinogen production in Komagataella phaffii through co-expression of HAC1 and PDI1.Microbial cell factories · 2026Article
- Systematic metabolic engineering of Komagataella phaffii for methanol-to-lactate bioconversion.Bioprocess and biosystems engineering · 2026Article
- Engineering serine metabolism to enhance AOX1 promoter self-induction in formate dehydrogenase-deficient Komagataella phaffii.Microbial cell factories · 2026Article
- Biological parts in yeast synthetic biology: from regulatory elements to predictive design platforms.FEMS yeast research · 2026Review
- Adaptive Laboratory Evolution of Pichia pastoris by Serial Batch Cultivation.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Design principle of successful genome editing applications using CRISPR-based toolkits.Journal of applied genetics · 2025Review
- Design of a Golden Gate Cloning Plasmid for the Generation of a Chimeric Virus-Like Particle-Based Subunit Vaccine Against Porcine Circovirus Type 2.Molecular biotechnology · 2025Article
- Sorbitol Uptake and Oxygen Transfer Shape AOX1 Promoter Induction in Formate Dehydrogenase-Deficient Komagataella phaffii.Microbial biotechnology · 2025Article
- Signaling pathways contributing to the transcriptional response to alkalinization in Komagataella phaffii: role of the transcription factors Crz1 and Rim101.Scientific reports · 2025Article
- Flo5-1 and Nrg1 are involved in reversible pH-dependent flocculation in Komagataella phaffii.Applied microbiology and biotechnology · 2025Article
- Illuminating Glucomannan Synthases To Explore Cell Wall Synthesis Bottlenecks.ACS synthetic biology · 2025Article
- Metabolic Engineering of Komagataella phaffii and Process Optimization for Biosynthesis of 1,2,4-Butanetriol From Xylose.Biotechnology journal · 2025Article
- A megatransposon drives the adaptation of Thermoanaerobacter kivui to carbon monoxide.Nature communications · 2025Article
- A toolkit for facilitating markerless integration of expression cassettes in Komagataella phaffii via CRISPR/Cas9.Microbial cell factories · 2025Article
- Expression and characterization of cold-adapted xylanase Xyl-L in Pichia pastoris for xylooligosaccharide (XOS) preparation.Microbial cell factories · 2025Article
- Metabolic engineering of Komagataella phaffii for enhanced 3-hydroxypropionic acid (3-HP) production from methanol.Journal of biological engineering · 2025Article
- Impact of Oxygen Availability on the Organelle-Specific Redox Potentials and Stress in Recombinant Protein Producing Komagataella phaffii.Microbial biotechnology · 2025Article
28 more citing papers are in PubMed but not listed here.
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Authors and funding
10 authors at 4 institutions in 2 countries.
Funding
Abstract
backgroundState-of-the-art strain engineering techniques for the host Pichia pastoris (syn. Komagataella spp.) include overexpression of homologous and heterologous genes, and deletion of host genes. For metabolic and cell engineering purposes the simultaneous overexpression of more than one gene would often be required. Very recently, Golden Gate based libraries were adapted to optimize single expression cassettes for recombinant proteins in P. pastoris. However, an efficient toolbox allowing the overexpression of multiple genes at once was not available for P. pastoris.
methodsWith the GoldenPiCS system, we provide a flexible modular system for advanced strain engineering in P. pastoris based on Golden Gate cloning. For this purpose, we established a wide variety of standardized genetic parts (20 promoters of different strength, 10 transcription terminators, 4 genome integration loci, 4 resistance marker cassettes).
resultsAll genetic parts were characterized based on their expression strength measured by eGFP as reporter in up to four production-relevant conditions. The promoters, which are either constitutive or regulatable, cover a broad range of expression strengths in their active conditions (2-192% of the glyceraldehyde-3-phosphate dehydrogenase promoter P
conclusionsGoldenPiCS, a modular Golden Gate-derived P. pastoris cloning system, is very flexible and efficient and can be used for strain engineering of P. pastoris to accomplish pathway expression, protein production or other applications where the integration of various DNA products is required. It allows for the assembly of up to eight expression units on one plasmid with the ability to use different characterized promoters and terminators for each expression unit. GoldenPiCS vectors are available at Addgene.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.