ArticleScience advances2025
A modular cell-free protein biosensor platform using split T7 RNA polymerase.
Article in Science advances, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Rapid-Forming Conductive Antifouling Hydrogel Coating Enables Magnetically Guided Electrochemical POCT for Direct at‑Home Blood Testing.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Biomolecular condensates can function as inherent catalysts.Molecular cell · 2026Article
- Microbial cell-free protein synthesis and its progression toward industrial use.Microbiology (Reading, England) · 2026Review
- A T7 RNAP regulatory toolbox for cell-free network engineering and biosensing applications.Nature communications · 2026Article
- Systematic Evaluation of Different Ribonucleoprotein Complexes as Posttranscriptional Biosensors in Cell-Free TX-TL Systems.ACS synthetic biology · 2026Article
- CF2H: a cell-free two-hybrid platform for rapid protein binder screening.Nature communications · 2026Article
- Programming Next-Generation Synthetic Biosensors by Genetic Circuit Design.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Review
- Computational redesign of a thermostable T7 RNA polymerase.Protein engineering, design & selection : PEDS · 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
- Computational redesign of a thermostable T7 RNA polymerase.bioRxiv : the preprint server for biology · 2025Article
- Harnessing Plasmid Crosstalk in Transcription Factor Mediated Cell-Free Biosensors.bioRxiv : the preprint server for biology · 2025Article
- Diversifying Substrates and Reaction Conditions for Polymerase Strand Recycling.ACS synthetic biology · 2025Article
- Recent Advances in Developing Cell-Free Protein Synthesis Biosensors for Medical Diagnostics and Environmental Monitoring.Biosensors · 2025Review
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Abstract
Conventional laboratory protein detection techniques are not suitable for point-of-care (POC) use because they require expensive equipment and laborious protocols, and existing POC assays suffer from long development timescales. Here, we describe a modular cell-free biosensing platform for generalizable protein detection that we call TLISA (T7 RNA polymerase-linked immunosensing assay), designed for extreme flexibility and equipment-free use. TLISA uses a split T7 RNA polymerase fused to affinity domains against a protein. The target antigen drives polymerase reassembly, inducing reporter expression. We characterize the platform and then demonstrate its modularity by using 16 affinity domains against four different antigens with minimal protocol optimization. We show that TLISA is suitable for POC use by sensing human biomarkers in serum and saliva with a colorimetric readout within 1 hour and by demonstrating functionality after lyophilization. Altogether, this technology has the potential to enable truly rapid, reconfigurable, modular, and equipment-free detection of diverse classes of proteins.
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