ArticleScientific reports2017
CASCADE, a platform for controlled gene amplification for high, tunable and selection-free gene expression in yeast.
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 7 papers.
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7 citing papers in PubMed, 20 citations in OpenAlex.
- Black yeasts are efficient heterologous hosts for the production of a wide range of fungal polyketides.Nature chemical biology · 2026Article
- RoCi-a single-step multi-copy integration system based on rolling-circle replication.Nucleic acids research · 2026Article
- Functional analysis of a rice 12-oxo-phytodienoic acid reductase gene (OsOPR1) involved in Cd stress tolerance.Molecular biology reports · 2024Article
- Comparison of Genome and Plasmid-Based Engineering of Multigene Benzylglucosinolate Pathway in Saccharomyces cerevisiae.Applied and environmental microbiology · 2022Article
- Integrating continuous hypermutation with high-throughput screening for optimization of cis,cis-muconic acid production in yeast.Microbial biotechnology · 2021Article
- Recent advances in the application of multiplex genome editing in Saccharomyces cerevisiae.Applied microbiology and biotechnology · 2021Review
- Evaluation of pyruvate decarboxylase-negativeEngineering in life sciences · 2019Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Over-expression of a gene by increasing its copy number is often desirable in the model yeast Saccharomyces cerevisiae. It may facilitate elucidation of enzyme functions, and in cell factory design it is used to increase production of proteins and metabolites. Current methods are typically exploiting expression from the multicopy 2 μ-derived plasmid or by targeting genes repeatedly into sequences like Ty or rDNA; in both cases, high gene expression levels are often reached. However, with 2 μ-based plasmid expression, the population of cells is very heterogeneous with respect to protein production; and for integration into repeated sequences it is difficult to determine the genetic setup of the resulting strains and to achieve specific gene doses. For both types of systems, the strains often suffer from genetic instability if proper selection pressure is not applied. Here we present a gene amplification system, CASCADE, which enables construction of strains with defined gene copy numbers. One or more genes can be amplified simultaneously and the resulting strains can be stably propagated on selection-free medium. As proof-of-concept, we have successfully used CASCADE to increase heterologous production of two fluorescent proteins, the enzyme β-galactosidase the fungal polyketide 6-methyl salicylic acid and the plant metabolite vanillin glucoside.
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