ArticleProtein engineering, design & selection : PEDS2014
SpeedyGenes: an improved gene synthesis method for the efficient production of error-corrected, synthetic protein libraries for directed evolution.
Article in Protein engineering, design & selection : PEDS, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 45 citations in OpenAlex.
- How far can you go? Extrapolating values of catalytic activity from known protein landscapes in natural and directed evolution.Chemical Society reviews · 2026Review
- Long oligodeoxynucleotides: chemical synthesis, isolation via catching-by-polymerization, verification via sequencing, and gene expression demonstration.Beilstein journal of organic chemistry · 2023Article
- Directed Evolution of Transcription Factor-Based Biosensors for Altered Effector Specificity.Methods in molecular biology (Clifton, N.J.) · 2022Article
- GeneORator: An Efficient Method for the Systematic Mutagenesis of Entire Genes.Methods in molecular biology (Clifton, N.J.) · 2022Article
- SpeedyGenesXL: an Automated, High-Throughput Platform for the Preparation of Bespoke Ultralarge Variant Libraries for Directed Evolution.Methods in molecular biology (Clifton, N.J.) · 2022Article
- A palette of fluorophores that are differentially accumulated by wild-type and mutant strains ofMicrobiology (Reading, England) · 2021Article
- GeneORator: An Effective Strategy for Navigating Protein Sequence Space More Efficiently through Boolean OR-Type DNA Libraries.ACS synthetic biology · 2019Article
- Experiment and Simulation Reveal How Mutations in Functional Plasticity Regions Guide Plant Monoterpene Synthase Product Outcome.ACS catalysis · 2019Article
- Directed evolution of the PcaV allosteric transcription factor to generate a biosensor for aromatic aldehydes.Journal of biological engineering · 2019Article
- Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting.The Analyst · 2018Article
- Engineering the "Missing Link" in Biosynthetic (-)-Menthol Production: Bacterial Isopulegone Isomerase.ACS catalysis · 2018Article
- Genetic tool development and systemic regulation in biosynthetic technology.Biotechnology for biofuels · 2018Review
- A systematic comparison of error correction enzymes by next-generation sequencing.Nucleic acids research · 2017Article
- Commentary on "Rapid identification of Streptococcus and Enterococcus species using diffuse reflectance-absorbance Fourier transform infrared spectroscopy and artificial neural networks".FEMS microbiology letters · 2017Article
- Evolutionary algorithms and synthetic biology for directed evolution: commentary on "on the mapping of genotype to phenotype in evolutionary algorithms" by Peter A. Whigham, Grant Dick, and James Maclaurin.Genetic programming and evolvable machines · 2017Article
- Computational tools for the evaluation of laboratory-engineered biocatalysts.Chemical communications (Cambridge, England) · 2016Review
- SYNBIOCHEM Synthetic Biology Research Centre, Manchester - A UK foundry for fine and speciality chemicals production.Synthetic and systems biotechnology · 2016Article
- Bioinformatics for the synthetic biology of natural products: integrating across the Design-Build-Test cycle.Natural product reports · 2016Review
- SYNBIOCHEM-a SynBio foundry for the biosynthesis and sustainable production of fine and speciality chemicals.Biochemical Society transactions · 2016Article
- Economical analysis of saturation mutagenesis experiments.Scientific reports · 2015Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
The de novo synthesis of genes is becoming increasingly common in synthetic biology studies. However, the inherent error rate (introduced by errors incurred during oligonucleotide synthesis) limits its use in synthesising protein libraries to only short genes. Here we introduce SpeedyGenes, a PCR-based method for the synthesis of diverse protein libraries that includes an error-correction procedure, enabling the efficient synthesis of large genes for use directly in functional screening. First, we demonstrate an accurate gene synthesis method by synthesising and directly screening (without pre-selection) a 747 bp gene for green fluorescent protein (yielding 85% fluorescent colonies) and a larger 1518 bp gene (a monoamine oxidase, producing 76% colonies with full catalytic activity, a 4-fold improvement over previous methods). Secondly, we show that SpeedyGenes can accommodate multiple and combinatorial variant sequences while maintaining efficient enzymatic error correction, which is particularly crucial for larger genes. In its first application for directed evolution, we demonstrate the use of SpeedyGenes in the synthesis and screening of large libraries of MAO-N variants. Using this method, libraries are synthesised, transformed and screened within 3 days. Importantly, as each mutation we introduce is controlled by the oligonucleotide sequence, SpeedyGenes enables the synthesis of large, diverse, yet controlled variant sequences for the purposes of directed evolution.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.