ArticleNucleic acids research2014
One-pot DNA construction for synthetic biology: the Modular Overlap-Directed Assembly with Linkers (MODAL) strategy.
Article in Nucleic acids research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 45 papers.
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Who cites it
45 citing papers in PubMed.
- Technologies for engineering repetitive DNA.Quantitative biology (Beijing, China) · 2026Review
- Advancing the frontier of plant-based therapeutics: critical innovations in molecular farming and bioprocess Integration.Biotechnology letters · 2026Review
- uSort-M: Scalable isolation of user-defined sequences from diverse pooled libraries.bioRxiv : the preprint server for biology · 2026Article
- Design of CRISPRi-Based Synthetic Gene Circuits in Bacteria.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Scaling DNA engineering.Trends in biotechnology · 2025Review
- Reusable combinatorial libraries with high diversity for efficient multi-gene expression optimization in Escherichia coli.World journal of microbiology & biotechnology · 2025Article
- CRISPR/Cas system-guided plasmid mutagenesis without sequence restriction.Fundamental research · 2025Article
- Methionyl-tRNA synthetase synthetic and proofreading activities are determinants of antibiotic persistence.Frontiers in microbiology · 2024Article
- Synthetic Gene Circuits Combining CRISPR Interference and CRISPR Activation inACS synthetic biology · 2023Article
- Article
- Functional Analysis of NovelMicroorganisms · 2023Article
- Robustness and innovation in synthetic genotype networks.Nature communications · 2023Article
- Evolution-aided engineering of plant specialized metabolism.aBIOTECH · 2021Review
- Reconstruction of Secondary Metabolic Pathway to Synthesize Novel Metabolite inFrontiers in bioengineering and biotechnology · 2021Article
- Recent Advances in Strategies for the Cloning of Natural Product Biosynthetic Gene Clusters.Frontiers in bioengineering and biotechnology · 2021Review
- Recent Advances in Silent Gene Cluster Activation inFrontiers in bioengineering and biotechnology · 2021Review
- Multistable and dynamic CRISPRi-based synthetic circuits.Nature communications · 2020Article
- Quantitative and Predictive Genetic Parts for Plant Synthetic Biology.Frontiers in plant science · 2020Review
- Rapid, modular, and cost-effective generation of donor DNA constructs for CRISPR-based gene knock-in.Biology methods & protocols · 2020Article
- Structural basis for the docking of mTORC1 on the lysosomal surface.Science (New York, N.Y.) · 2019Article
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7 authors.
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Abstract
Overlap-directed DNA assembly methods allow multiple DNA parts to be assembled together in one reaction. These methods, which rely on sequence homology between the ends of DNA parts, have become widely adopted in synthetic biology, despite being incompatible with a key principle of engineering: modularity. To answer this, we present MODAL: a Modular Overlap-Directed Assembly with Linkers strategy that brings modularity to overlap-directed methods, allowing assembly of an initial set of DNA parts into a variety of arrangements in one-pot reactions. MODAL is accompanied by a custom software tool that designs overlap linkers to guide assembly, allowing parts to be assembled in any specified order and orientation. The in silico design of synthetic orthogonal overlapping junctions allows for much greater efficiency in DNA assembly for a variety of different methods compared with using non-designed sequence. In tests with three different assembly technologies, the MODAL strategy gives assembly of both yeast and bacterial plasmids, composed of up to five DNA parts in the kilobase range with efficiencies of between 75 and 100%. It also seamlessly allows mutagenesis to be performed on any specified DNA parts during the process, allowing the one-step creation of construct libraries valuable for synthetic biology applications.
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