ArticleNucleic acids research2016
Integrating gene synthesis and microfluidic protein analysis for rapid protein engineering.
Article in Nucleic acids research, 2016. 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.
- A T7 RNAP regulatory toolbox for cell-free network engineering and biosensing applications.Nature communications · 2026Article
- Cell-free immuno-profiling on a genetically programmed biochip.Nature nanotechnology · 2026Article
- Regulatory Components for Bacterial Cell-Free Systems Engineering.ACS synthetic biology · 2024Review
- Rugged fitness landscapes minimize promiscuity in the evolution of transcriptional repressors.Cell systems · 2024Article
- Systematic analysis of low-affinity transcription factor binding site clusters in vitro and in vivo establishes their functional relevance.Nature communications · 2022Article
- CFPU: A Cell-Free Processing Unit for High-Throughput, Automated In Vitro Circuit Characterization in Steady-State Conditions.Biodesign research · 2021Article
- A partially self-regenerating synthetic cell.Nature communications · 2020Article
- Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology.Frontiers in bioengineering and biotechnology · 2020Review
- Cell-free gene-regulatory network engineering with synthetic transcription factors.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Hybrid cell reactor system from Escherichia coli protoplast cells and arrayed lipid bilayer chamber device.Scientific reports · 2018Article
- Integrating Immunology and Microfluidics for Single Immune Cell Analysis.Frontiers in immunology · 2018Review
- Novel methods to optimize gene and statistic test for evaluation - an application for Escherichia coli.BMC bioinformatics · 2017Article
- Engineering Translation in Mammalian Cell Factories to Increase Protein Yield: The Unexpected Use of Long Non-Coding SINEUP RNAs.Computational and structural biotechnology journal · 2016Review
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Authors and funding
4 authors.
Funding
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Abstract
The capability to rapidly design proteins with novel functions will have a significant impact on medicine, biotechnology and synthetic biology. Synthetic genes are becoming a commodity, but integrated approaches have yet to be developed that take full advantage of gene synthesis. We developed a solid-phase gene synthesis method based on asymmetric primer extension (APE) and coupled this process directly to high-throughput, on-chip protein expression, purification and characterization (via mechanically induced trapping of molecular interactions, MITOMI). By completely circumventing molecular cloning and cell-based steps, APE-MITOMI reduces the time between protein design and quantitative characterization to 3-4 days. With APE-MITOMI we synthesized and characterized over 400 zinc-finger (ZF) transcription factors (TF), showing that although ZF TFs can be readily engineered to recognize a particular DNA sequence, engineering the precise binding energy landscape remains challenging. We also found that it is possible to engineer ZF-DNA affinity precisely and independently of sequence specificity and that in silico modeling can explain some of the observed affinity differences. APE-MITOMI is a generic approach that should facilitate fundamental studies in protein biophysics, and protein design/engineering.
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