ArticleACS synthetic biology2012
A Heritable Recombination system for synthetic Darwinian evolution in yeast.
Article in ACS synthetic biology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Evolution and synthetic biology.Current opinion in microbiology · 2023Review
- Synthetic biology: at the crossroads of genetic engineering and human therapeutics-a Keystone Symposia report.Annals of the New York Academy of Sciences · 2021Review
- Towards an engineering theory of evolution.Nature communications · 2021Review
- The developing toolkit of continuous directed evolution.Nature chemical biology · 2020Review
- Probing pathways of adaptation with continuous evolution.Current opinion in systems biology · 2019Article
- In vivo biosensors: mechanisms, development, and applications.Journal of industrial microbiology & biotechnology · 2018Article
- Modern methods for laboratory diversification of biomolecules.Current opinion in chemical biology · 2017Review
- Recurrent RNA motifs as scaffolds for genetically encodable small-molecule biosensors.Nature chemical biology · 2017Article
- Targeted AID-mediated mutagenesis (TAM) enables efficient genomic diversification in mammalian cells.Nature methods · 2016Article
- In vivo continuous evolution of genes and pathways in yeast.Nature communications · 2016Article
- Direct Capture Technologies for Genomics-Guided Discovery of Natural Products.Current topics in medicinal chemistry · 2016Review
- Methods for the directed evolution of proteins.Nature reviews. Genetics · 2015Review
- In vivo continuous directed evolution.Current opinion in chemical biology · 2015Review
- Molecular tools for chemical biotechnology.Current opinion in biotechnology · 2013Review
- Designing de novo: interdisciplinary debates in synthetic biology.Systems and synthetic biology · 2013Article
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Authors and funding
4 authors.
Funding
Abstract
Genetic recombination is central to the generation of molecular diversity and enhancement of evolutionary fitness in living systems. Methods such as DNA shuffling that recapitulate this diversity mechanism in vitro are powerful tools for engineering biomolecules with useful new functions by directed evolution. Synthetic biology now brings demand for analogous technologies that enable the controlled recombination of beneficial mutations in living cells. Thus, here we create a Heritable Recombination system centered around a library cassette plasmid that enables inducible mutagenesis via homologous recombination and subsequent combination of beneficial mutations through sexual reproduction in Saccharomyces cerevisiae. Using repair of nonsense codons in auxotrophic markers as a model, Heritable Recombination was optimized to give mutagenesis efficiencies of up to 6% and to allow successive repair of different markers through two cycles of sexual reproduction and recombination. Finally, Heritable Recombination was employed to change the substrate specificity of a biosynthetic enzyme, with beneficial mutations in three different active site loops crossed over three continuous rounds of mutation and selection to cover a total sequence diversity of 10(13). Heritable Recombination, while at an early stage of development, breaks the transformation barrier to library size and can be immediately applied to combinatorial crossing of beneficial mutations for cell engineering, adding important features to the growing arsenal of next generation molecular biology tools for synthetic biology.
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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.