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• W3178149625 abstract "Abstract The new generation of cell-free gene expression systems enables the prototyping and engineering of biological systems in vitro over a remarkable scope of applications and physical scales. As the utilization of DNA-directed in vitro protein synthesis expands in scope, developing more powerful cell-free transcription–translation (TXTL) platforms remains a major goal to either execute larger DNA programs or improve cell-free biomanufacturing capabilities. In this work, we report the capabilities of the all-E. coli TXTL toolbox 3.0, a multipurpose cell-free expression system specifically developed for synthetic biology. In non-fed batch-mode reactions, the synthesis of the fluorescent reporter protein eGFP (enhanced green fluorescent protein) reaches 4 mg/ml. In synthetic cells, consisting of liposomes loaded with a TXTL reaction, eGFP is produced at concentrations of &gt;8 mg/ml when the chemical building blocks feeding the reaction diffuse through membrane channels to facilitate exchanges with the outer solution. The bacteriophage T7, encoded by a genome of 40 kb and ∼60 genes, is produced at a concentration of 1013 PFU/ml (plaque forming unit/ml). This TXTL system extends the current cell-free expression capabilities by offering unique strength and properties, for testing regulatory elements and circuits, biomanufacturing biologics or building synthetic cells." @default.
• W3178149625 created "2021-07-19" @default.
• W3178149625 creator A5048707902 @default.
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• W3178149625 creator A5083430931 @default.
• W3178149625 date "2021-08-04" @default.
• W3178149625 modified "2023-10-16" @default.
• W3178149625 title "The all-E. coliTXTL toolbox 3.0: new capabilities of a cell-free synthetic biology platform" @default.
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• W3178149625 doi "https://doi.org/10.1093/synbio/ysab017" @default.
• W3178149625 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/8546610" @default.
• W3178149625 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34712841" @default.
• W3178149625 hasPublicationYear "2021" @default.
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