Reference - Detail
| RRC ID | 86546 |
|---|---|
| Author | Tominaga M, Nozaki K, Umeno D, Ishii J, Kondo A. |
| Title | Robust and flexible platform for directed evolution of yeast genetic switches. |
| Journal | Nat Commun |
| Abstract |
A wide repertoire of genetic switches has accelerated prokaryotic synthetic biology, while eukaryotic synthetic biology has lagged in the model organism Saccharomyces cerevisiae. Eukaryotic genetic switches are larger and more complex than prokaryotic ones, complicating the rational design and evolution of them. Here, we present a robust workflow for the creation and evolution of yeast genetic switches. The selector system was designed so that both ON- and OFF-state selection of genetic switches is completed solely by liquid handling, and it enabled parallel screen/selection of different motifs with different selection conditions. Because selection threshold of both ON- and OFF-state selection can be flexibly tuned, the desired selection conditions can be rapidly pinned down for individual directed evolution experiments without a prior knowledge either on the library population. The system's utility was demonstrated using 20 independent directed evolution experiments, yielding genetic switches with elevated inducer sensitivities, inverted switching behaviours, sensory functions, and improved signal-to-noise ratio (>100-fold induction). The resulting yeast genetic switches were readily integrated, in a plug-and-play manner, into an AND-gated carotenoid biosynthesis pathway. |
| Volume | 12(1) |
| Pages | 1846 |
| Published | 2021-3-23 |
| DOI | 10.1038/s41467-021-22134-y |
| PII | 10.1038/s41467-021-22134-y |
| PMID | 33758180 |
| PMC | PMC7988172 |
| MeSH | Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism Basidiomycota / genetics Basidiomycota / metabolism Directed Molecular Evolution / methods* Flow Cytometry Gene Library Genes, Reporter Genes, Switch* Genetic Engineering / methods* Genetic Techniques* Phloroglucinol / analogs & derivatives Phloroglucinol / pharmacology Promoter Regions, Genetic Repressor Proteins / chemistry Repressor Proteins / genetics Repressor Proteins / metabolism Saccharomyces cerevisiae / genetics* Signal-To-Noise Ratio Synthetic Biology / methods* Tetracycline / pharmacology Trans-Activators / chemistry Trans-Activators / genetics Trans-Activators / metabolism beta Carotene / biosynthesis beta Carotene / genetics beta Carotene / metabolism |
| IF | 12.121 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 25 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| DNA material | pGK416m-pphlO6-ymUkG1 (RDB21032) |