ArticleNature chemical biology2023
Engineered bacterial orthogonal DNA replication system for continuous evolution.
Article in Nature chemical biology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed, 57 citations in OpenAlex.
- Harnessing the central dogma: Enzyme-driven gene diversification strategies for in cellulo continuous directed evolution.Fundamental research · 2026Review
- Continuous Evolution System Based on Toxin-Antitoxin System Combined with Base Deaminase Accelerates the In Vivo Modification of Target Proteins.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Exploring the Evolutionary Landscape with Targeted In Vivo Hypermutations.Biomedicines · 2026Review
- Replication-coupled dual-deaminase mutagenesis (RCDM) enables continuous genomic evolution in Bacillus subtilis.Microbial cell factories · 2026Article
- Construction of synthetic protein-binding non-genetic DNA systems in living cells.Nature chemistry · 2026Article
- Bridging continuous and discrete evolution through a controllable, hypermutagenic phage-bacteria system.Nature microbiology · 2026Article
- Diversity-generating retroelements for programmable targeted hypermutagenesis.Nature biotechnology · 2026Article
- MicroDFBEST: A dCas12b-derived dual-function base editor with programmable editing characteristics for microbial genetic engineering.Synthetic and systems biotechnology · 2026Article
- Highly mutagenic continuous evolution in E. coli using a Φ29-based orthogonal replication system.Nature biotechnology · 2026Article
- Design prokaryotic cis-regulatory elements using language model.Nucleic acids research · 2026Article
- Alternative nucleic acid structures in microbial biology: regulatory functions and antimicrobial opportunities.Frontiers in cellular and infection microbiology · 2026Review
- Glycoengineering strategies for constructing defined Mucin O-glycans.Frontiers in molecular biosciences · 2026Review
- CRISPR-DNA Polymerase Assisted Targeted Mutagenesis for Regulable Laboratory Evolution.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- A super protein evolution engine.Nature chemical biology · 2025Article
- Editor's Choice Protein engineering strategies to develop lectins by design.Glycobiology · 2025Review
- T7 RNA polymerase-guided base editor for accelerated continuous evolution inSynthetic and systems biotechnology · 2025Article
- Combing Directed Enzyme Evolution with Metabolic Engineering to Develop Efficient Microbial Cell Factories.Chem & bio engineering · 2025Review
- Considering Metabolic Context in Enzyme Evolution and Design.Biochemistry · 2025Review
- An orthogonal transcription mutation system generating all transition mutations for accelerated protein evolution in vivo.Nature communications · 2025Article
- Ultra-efficient Integration of Gene Libraries onto Yeast Cytosolic Plasmids.ACS synthetic biology · 2025Article
Corrections and comments
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Authors and funding
12 authors at 1 institution in 1 country.
Funding
Abstract
Continuous evolution can generate biomolecules for synthetic biology and enable evolutionary investigation. The orthogonal DNA replication system (OrthoRep) in yeast can efficiently mutate long DNA fragments in an easy-to-operate manner. However, such a system is lacking in bacteria. Therefore, we developed a bacterial orthogonal DNA replication system (BacORep) for continuous evolution. We achieved this by harnessing the temperate phage GIL16 DNA replication machinery in Bacillus thuringiensis with an engineered error-prone orthogonal DNA polymerase. BacORep introduces all 12 types of nucleotide substitution in 15-kilobase genes on orthogonally replicating linear plasmids with a 6,700-fold higher mutation rate than that of the host genome, the mutation rate of which is unchanged. Here we demonstrate the utility of BacORep-based continuous evolution by generating strong promoters applicable to model bacteria, Bacillus subtilis and Escherichia coli, and achieving a 7.4-fold methanol assimilation increase in B. thuringiensis. BacORep is a powerful tool for continuous evolution in prokaryotic cells.
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Registered trials
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.