ArticleNature methods2023
Fast and efficient template-mediated synthesis of genetic variants.
Article in Nature methods, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
What it found
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Who cites it
10 citing papers in PubMed, 18 citations in OpenAlex.
- High-throughput synthesis of DNA fragments by molecular self-assembly of overlapping oligonucleotides.Nature biotechnology · 2026Article
- Rapid directed evolution guided by protein language models and epistatic interactions.Science (New York, N.Y.) · 2026Article
- Accelerating primer design for amplicon sequencing using large language model-powered agents.Nature biomedical engineering · 2026Article
- From liquid-phase synthesis to chemical ligation: preparation of oligonucleotides and their backbone analogs in solution.Nucleic acids research · 2025Review
- The design and engineering of synthetic genomes.Nature reviews. Genetics · 2025Review
- Oligonucleotide subsets selection by single nucleotide resolution barcode identification.Nature communications · 2025Article
- Hybrid Exb/Mot stators require substitutions distant from the chimeric pore to power flagellar rotation.Journal of bacteriology · 2024Article
- Template-dependent DNA ligation for the synthesis of modified oligonucleotides.Nature communications · 2024Article
- Controlled enzymatic synthesis of oligonucleotides.Communications chemistry · 2024Review
- Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Efficient methods for the generation of specific mutations enable the study of functional variations in natural populations and lead to advances in genetic engineering applications. Here, we present a new approach, mutagenesis by template-guided amplicon assembly (MEGAA), for the rapid construction of kilobase-sized DNA variants. With this method, many mutations can be generated at a time to a DNA template at more than 90% efficiency per target in a predictable manner. We devised a robust and iterative protocol for an open-source laboratory automation robot that enables desktop production and long-read sequencing validation of variants. Using this system, we demonstrated the construction of 31 natural SARS-CoV2 spike gene variants and 10 recoded Escherichia coli genome fragments, with each 4 kb region containing up to 150 mutations. Furthermore, 125 defined combinatorial adeno-associated virus-2 cap gene variants were easily built using the system, which exhibited viral packaging enhancements of up to 10-fold compared with wild type. Thus, the MEGAA platform enables generation of multi-site sequence variants quickly, cheaply, and in a scalable manner for diverse applications in biotechnology.
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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.