ArticleACS synthetic biology2024
An Automated Cell-Free Workflow for Transcription Factor Engineering.
Article in ACS synthetic biology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Microbial cell-free protein synthesis and its progression toward industrial use.Microbiology (Reading, England) · 2026Review
- A liquid handling platform for standardized quantification of cell-free enzymatic activity encoded by antimicrobial resistance genes.Synthetic biology (Oxford, England) · 2026Article
- Active learning-guided optimization of cell-free biosensors for lead testing in drinking water.Nature communications · 2025Article
- Whole-cell and cell-free biosensor-driven metabolic engineering.Current opinion in biotechnology · 2025Review
- An AI-driven workflow for the accelerated optimization of cell-free protein synthesis.iScience · 2025Article
- Reconstituting alternative life using the test-bed of cell-free systems.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2025Review
- Challenges and Opportunities in Smart Biosensing for Biomanufacturing.ACS synthetic biology · 2025Review
- Automated and Programmable Cell-Free Systems for Scalable Synthetic Biology with a Focus on Biofoundry Integration.Journal of microbiology and biotechnology · 2025Review
- One-pot cloning and protein expression platform for genetic engineering.bioRxiv : the preprint server for biology · 2025Article
- Characterizing and engineering post-translational modifications with high-throughput cell-free expression.Nature communications · 2025Article
- Cell-Free Protein Synthesis as a Method to Rapidly Screen Machine Learning-Generated Protease Variants.ACS synthetic biology · 2025Article
- Developing, Characterizing, and Modeling CRISPR-Based Point-of-Use Pathogen Diagnostics.ACS synthetic biology · 2025Article
- Regulatory Components for Bacterial Cell-Free Systems Engineering.ACS synthetic biology · 2024Review
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Authors and funding
6 authors.
Funding
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Abstract
The design and optimization of metabolic pathways, genetic systems, and engineered proteins rely on high-throughput assays to streamline design-build-test-learn cycles. However, assay development is a time-consuming and laborious process. Here, we create a generalizable approach for the tailored optimization of automated cell-free gene expression (CFE)-based workflows, which offers distinct advantages over in vivo assays in reaction flexibility, control, and time to data. Centered around designing highly accurate and precise transfers on the Echo Acoustic Liquid Handler, we introduce pilot assays and validation strategies for each stage of protocol development. We then demonstrate the efficacy of our platform by engineering transcription factor-based biosensors. As a model, we rapidly generate and assay libraries of 127 MerR and 134 CadR transcription factor variants in 3682 unique CFE reactions in less than 48 h to improve limit of detection, selectivity, and dynamic range for mercury and cadmium detection. This was achieved by assessing a panel of ligand conditions for sensitivity (to 0.1, 1, 10 μM Hg and 0, 1, 10, 100 μM Cd for MerR and CadR, respectively) and selectivity (against Ag, As, Cd, Co, Cu, Hg, Ni, Pb, and Zn). We anticipate that our Echo-based, cell-free approach can be used to accelerate multiple design workflows in synthetic biology.
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Registered trials
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