ArticleNucleic acids research2021
Gene-specific mutagenesis enables rapid continuous evolution of enzymes in vivo.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 41 papers.
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41 citing papers in PubMed, 82 citations in OpenAlex.
- Coupling continuous directed evolution with base editors identifies combinatorial antibiotic resistance.Biotechnology letters · 2026Article
- Harnessing the central dogma: Enzyme-driven gene diversification strategies for in cellulo continuous directed evolution.Fundamental research · 2026Review
- Sequence-free landscape inference for directed evolution.PLoS computational biology · 2026Article
- Exploring the Evolutionary Landscape with Targeted In Vivo Hypermutations.Biomedicines · 2026Review
- Base Editors for Engineering Industrial Microorganisms: Types, Applications, and Future Perspectives.Biotechnology journal · 2026Review
- Engineering T7 RNA polymerase-cascaded systems controlled by nisin and theophylline for protein overexpression and targeted gene mutagenesis inSynthetic and systems biotechnology · 2025Article
- Continuous targeted hypermutation with a tunable mutation window.Nature communications · 2025Article
- T7 RNA polymerase-guided base editor for accelerated continuous evolution inSynthetic and systems biotechnology · 2025Article
- Modification and applications of glucose oxidase: optimization strategies and high-throughput screening technologies.World journal of microbiology & biotechnology · 2025Review
- In vivo directed evolution of an ultrafast Rubisco from a semianaerobic environment imparts oxygen resistance.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- An orthogonal transcription mutation system generating all transition mutations for accelerated protein evolution in vivo.Nature communications · 2025Article
- A chimeric viral platform for directed evolution in mammalian cells.Nature communications · 2025Article
- Growth-coupled continuous directed evolution by MutaT7 enables efficient and automated enzyme engineering.Applied and environmental microbiology · 2025Article
- Recent advances in targeted mutagenesis to expedite the evolution of biological systems.Journal of microbiology (Seoul, Korea) · 2025Review
- A Golden Gate compatible system for continuous directed evolution inSynthetic biology (Oxford, England) · 2025Article
- Targeted mutagenesis of specific genomic DNA sequences in animals for thebioRxiv : the preprint server for biology · 2024Article
- New-to-nature CONature communications · 2024Article
- MutaT7ACS synthetic biology · 2024Article
- Engineering is evolution: a perspective on design processes to engineer biology.Nature communications · 2024Article
- Automated in vivo enzyme engineering accelerates biocatalyst optimization.Nature communications · 2024Review
Corrections and comments
- Erratum issued
Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Various in vivo mutagenesis methods have been developed to facilitate fast and efficient continuous evolution of proteins in cells. However, they either modify the DNA region that does not match the target gene, or suffer from low mutation rates. Here, we report a mutator, eMutaT7 (enhanced MutaT7), with very fast in vivo mutation rate and high gene-specificity in Escherichia coli. eMutaT7, a cytidine deaminase fused to an orthogonal RNA polymerase, can introduce up to ∼4 mutations per 1 kb per day, rivalling the rate in typical in vitro mutagenesis for directed evolution of proteins, and promotes rapid continuous evolution of model proteins for antibiotic resistance and allosteric activation. eMutaT7 provides a very simple and tunable method for continuous directed evolution of proteins, and suggests that the fusion of new DNA-modifying enzymes to the orthogonal RNA polymerase is a promising strategy to explore the expanded sequence space without compromising gene specificity.
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