ArticleACS synthetic biology2018
Tunable Expression Systems for Orthogonal DNA Replication.
Article in ACS synthetic biology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Continuous Hypermutation and Evolution of Luciferase Variants.ACS chemical biology · 2026Article
- Article
- Mirages in continuous directed enzyme evolution: a cautionary case study with plantized bacterial THI4 enzymes.Plant biotechnology journal · 2025Article
- Recent advances in targeted mutagenesis to expedite the evolution of biological systems.Journal of microbiology (Seoul, Korea) · 2025Review
- Harnessing mass spectrometry-based proteomics for continuous directed evolution.Synthetic biology (Oxford, England) · 2025Article
- Continuous evolution of user-defined genes at 1 million times the genomic mutation rate.Science (New York, N.Y.) · 2024Article
- Adapting enzymes to improve their functionality in plants: why and how.Biochemical Society transactions · 2023Review
- Using continuous directed evolution to improve enzymes for plant applications.Plant physiology · 2022Article
- In vivo hypermutation and continuous evolution.Nature reviews. Methods primers · 2022Article
- Systems for in vivo hypermutation: a quest for scale and depth in directed evolution.Current opinion in chemical biology · 2021Review
- Rapid generation of potent antibodies by autonomous hypermutation in yeast.Nature chemical biology · 2021Article
- Genome editor-directed in vivo library diversification.Cell chemical biology · 2021Review
- Winner-takes-all resource competition redirects cascading cell fate transitions.Nature communications · 2021Article
- Phage-assisted continuous and non-continuous evolution.Nature protocols · 2020Article
- Scalable continuous evolution for the generation of diverse enzyme variants encompassing promiscuous activities.Nature communications · 2020Article
- Potential for Applying Continuous Directed Evolution to Plant Enzymes: An Exploratory Study.Life (Basel, Switzerland) · 2020Article
- Topology-dependent interference of synthetic gene circuit function by growth feedback.Nature chemical biology · 2020Article
- Messenger RNAs of Yeast Virus-Like Elements Contain Non-templated 5' Poly(A) Leaders, and Their Expression Is Independent of eIF4E and Pab1.Frontiers in microbiology · 2019Article
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3 authors.
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Abstract
We recently developed an orthogonal DNA replication (OrthoRep) system capable of driving the rapid continuous evolution of genes in vivo. However, OrthoRep uses a special transcription system, the components of which (e.g., promoters) have previously limited the strength with which OrthoRep-encoded genes can be expressed. Here, we report a collection of synthetic and evolved OrthoRep expression parts that allow OrthoRep-encoded genes to span expression levels matching those of endogenous Saccharomyces cerevisiae genes. Specifically, we found that various promoter mutations as well as a genetically encoded poly(A) tail enable us to tune the expression level of OrthoRep-encoded genes over a large range and up to levels 43-fold higher than were previously attained, reaching at least ∼40% of the strength of the genomic TDH3 promoter. We further show that expression level gains using our new parts are stable over passaging and consistent across multiple genes and OrthoRep systems of different mutation rates. This new set of expression parts further expands OrthoRep's applicability to the continuous in vivo evolution of proteins and pathways.
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