ArticleNucleic acids research2018
MetClo: methylase-assisted hierarchical DNA assembly using a single type IIS restriction enzyme.
Article in Nucleic acids research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- The plant Golden Gate toolkit: compatibility, interoperability, and harmonized assembly in plant synthetic biology.Plant methods · 2026Review
- UniClo technology exploits methylation for universal scarless DNA assembly.Biology methods & protocols · 2026Review
- Construction and Diversification of Natural Product Biosynthetic Gene Clusters at High Efficiency and Accuracy.ACS synthetic biology · 2025Article
- Novel Type IIS-Based Library Assembly Technique for Developing Nanobodies Targeting IPNv VP2 Protein.International journal of molecular sciences · 2025Article
- DASH: a versatile and high-capacity gene stacking system for plant synthetic biology.Plant biotechnology journal · 2025Review
- UniClo: scarless hierarchical DNA assembly without sequence constraint.Nucleic acids research · 2025Article
- Synthetic Biology in Natural Product Biosynthesis.Chemical reviews · 2025Review
- Golden Gate Cloning of Multigene Constructs Using the Modular Cloning System MoClo.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Multivariate modular metabolic engineering and medium optimization for vitamin BSynthetic and systems biotechnology · 2024Article
- Directed Evolution of Acoustic Reporter Genes Using High-Throughput Acoustic Screening.ACS synthetic biology · 2024Article
- DNA methylases for site-selective inhibition of type IIS restriction enzyme activity.Applied microbiology and biotechnology · 2024Article
- Rapid 40 kb Genome Construction from 52 Parts through Data-optimized Assembly Design.ACS synthetic biology · 2022Article
- Having a Same Type IIS Enzyme's Restriction Site on Guide RNA Sequence Does Not Affect Golden Gate (GG) Cloning and Subsequent CRISPR/Cas Mutagenesis.International journal of molecular sciences · 2022Article
- Computer-Aided Design and Pre-validation of Large Batches of DNA Assemblies.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Universal loop assembly: open, efficient and cross-kingdom DNA fabrication.Synthetic biology (Oxford, England) · 2020Article
- Article
- Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design.PloS one · 2020Article
- Nimble Cloning: A Simple, Versatile, and Efficient System for Standardized Molecular Cloning.Frontiers in bioengineering and biotechnology · 2019Article
- Highly efficient construction of infectious viroid-derived clones.Plant methods · 2019Article
Corrections and comments
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Authors and funding
2 authors.
Funding
Abstract
Efficient DNA assembly is of great value in biological research and biotechnology. Type IIS restriction enzyme-based assembly systems allow assembly of multiple DNA fragments in a one-pot reaction. However, large DNA fragments can only be assembled by alternating use of two or more type IIS restriction enzymes in a multi-step approach. Here, we present MetClo, a DNA assembly method that uses only a single type IIS restriction enzyme for hierarchical DNA assembly. The method is based on in vivo methylation-mediated on/off switching of type IIS restriction enzyme recognition sites that overlap with site-specific methylase recognition sequences. We have developed practical MetClo systems for the type IIS enzymes BsaI, BpiI and LguI, and demonstrated hierarchical assembly of large DNA fragments up to 218 kb. The MetClo approach substantially reduces the need to remove internal restriction sites from components to be assembled. The use of a single type IIS enzyme throughout the different stages of DNA assembly allows novel and powerful design schemes for rapid large-scale hierarchical DNA assembly. The BsaI-based MetClo system is backward-compatible with component libraries of most of the existing type IIS restriction enzyme-based assembly systems, and has potential to become a standard for modular DNA assembly.
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Registered trials
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