ArticleNature communications2021
An integrated in vivo/in vitro framework to enhance cell-free biosynthesis with metabolically rewired yeast extracts.
Article in Nature communications, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 35 citations in OpenAlex.
- Review
- Eukaryotic cell-free protein synthesis: Chassis diversification, system engineering, and emerging applications.Synthetic and systems biotechnology · 2026Review
- Microbial cell-free protein synthesis and its progression toward industrial use.Microbiology (Reading, England) · 2026Review
- Advances in natural product discovery: strategies, technologies, and insights.Natural products and bioprospecting · 2026Review
- Automated and Programmable Cell-Free Systems for Scalable Synthetic Biology with a Focus on Biofoundry Integration.Journal of microbiology and biotechnology · 2025Review
- Cell-Free Systems to Mimic and Expand Metabolism.ACS synthetic biology · 2025Review
- BeyondChemical reviews · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Review
- Shifting redox reaction equilibria on demand using an orthogonal redox cofactor.Nature chemical biology · 2024Article
- Chloroplast Cell-Free Systems from Different Plant Species as a Rapid Prototyping Platform.ACS synthetic biology · 2024Article
- Usage of Cell-Free Protein Synthesis in Post-Translational Modification of μ-Conopeptide PIIIA.Marine drugs · 2023Article
- A Low-Cost, Thermostable, Cell-Free Protein Synthesis Platform for On-Demand Production of Conjugate Vaccines.ACS synthetic biology · 2023Article
- Rewiring cell-free metabolic flux inSynthetic biology (Oxford, England) · 2023Article
- Cell-Free Display Techniques for Protein Evolution.Advances in biochemical engineering/biotechnology · 2023Article
- Alternative design strategies to help build the enzymatic retrosynthesis toolbox.RSC chemical biology · 2022Article
- Engineering Embden-Meyerhof-Parnas Glycolysis to Generate Noncanonical Reducing Power.ACS catalysis · 2022Article
- Systems biology-based analysis of cell-free systems.Current opinion in biotechnology · 2022Review
- Cell-free prototyping enables implementation of optimized reverse β-oxidation pathways in heterotrophic and autotrophic bacteria.Nature communications · 2022Article
Corrections and comments
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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
Cell-free systems using crude cell extracts present appealing opportunities for designing biosynthetic pathways and enabling sustainable chemical synthesis. However, the lack of tools to effectively manipulate the underlying host metabolism in vitro limits the potential of these systems. Here, we create an integrated framework to address this gap that leverages cell extracts from host strains genetically rewired by multiplexed CRISPR-dCas9 modulation and other metabolic engineering techniques. As a model, we explore conversion of glucose to 2,3-butanediol in extracts from flux-enhanced Saccharomyces cerevisiae strains. We show that cellular flux rewiring in several strains of S. cerevisiae combined with systematic optimization of the cell-free reaction environment significantly increases 2,3-butanediol titers and volumetric productivities, reaching productivities greater than 0.9 g/L-h. We then show the generalizability of the framework by improving cell-free itaconic acid and glycerol biosynthesis. Our coupled in vivo/in vitro metabolic engineering approach opens opportunities for synthetic biology prototyping efforts and cell-free biomanufacturing.
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Registered trials
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