ArticleProceedings of the National Academy of Sciences of the United States of America2011
Reiterative Recombination for the in vivo assembly of libraries of multigene pathways.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed, 128 citations in OpenAlex.
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- Precise Editing at DNA Replication Forks Enables Multiplex Genome Engineering in Eukaryotes.Cell · 2017Article
- High-throughput characterization of protein-protein interactions by reprogramming yeast mating.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- A modular yeast biosensor for low-cost point-of-care pathogen detection.Science advances · 2017Article
- Technology development for natural product biosynthesis in Saccharomyces cerevisiae.Current opinion in biotechnology · 2016Review
- Engineering microbial hosts for production of bacterial natural products.Natural product reports · 2016Review
- EasyClone-MarkerFree: A vector toolkit for marker-less integration of genes into Saccharomyces cerevisiae via CRISPR-Cas9.Biotechnology journal · 2016Article
- New tools for reconstruction and heterologous expression of natural product biosynthetic gene clusters.Natural product reports · 2016Review
- A Barcoding Strategy Enabling Higher-Throughput Library Screening by Microscopy.ACS synthetic biology · 2015Article
- A tetO Toolkit To Alter Expression of Genes in Saccharomyces cerevisiae.ACS synthetic biology · 2015Article
- Recent advances in DNA assembly technologies.FEMS yeast research · 2015Review
- EMBO conference series: Chemical Biology 2014.Chembiochem : a European journal of chemical biology · 2014Article
- A system for multilocus chromosomal integration and transformation-free selection marker rescue.FEMS yeast research · 2014Article
- Synthetic biology and metabolic engineering for marine carotenoids: new opportunities and future prospects.Marine drugs · 2014Review
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2 authors at 1 institution in 1 country.
Funding
Abstract
The increasing sophistication of synthetic biology is creating a demand for robust, broadly accessible methodology for constructing multigene pathways inside of the cell. Due to the difficulty of rationally designing pathways that function as desired in vivo, there is a further need to assemble libraries of pathways in parallel, in order to facilitate the combinatorial optimization of performance. While some in vitro DNA assembly methods can theoretically make libraries of pathways, these techniques are resource intensive and inherently require additional techniques to move the DNA back into cells. All previously reported in vivo assembly techniques have been low yielding, generating only tens to hundreds of constructs at a time. Here, we develop "Reiterative Recombination," a robust method for building multigene pathways directly in the yeast chromosome. Due to its use of endonuclease-induced homologous recombination in conjunction with recyclable markers, Reiterative Recombination provides a highly efficient, technically simple strategy for sequentially assembling an indefinite number of DNA constructs at a defined locus. In this work, we describe the design and construction of the first Reiterative Recombination system in Saccharomyces cerevisiae, and we show that it can be used to assemble multigene constructs. We further demonstrate that Reiterative Recombination can construct large mock libraries of at least 10(4) biosynthetic pathways. We anticipate that our system's simplicity and high efficiency will make it a broadly accessible technology for pathway construction and render it a valuable tool for optimizing pathways in vivo.
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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.