A robust gene-stacking method utilizing yeast assembly for plant synthetic biology

• DOI: 10.1038/ncomms13215
• Publication Date: 2016-10-26T11:22:26Z
• Authors: Patrick M. Shih; Khanh Vuu; Nasim Mansoori; Leïla Ayad; Katherine B. Louie; Benjamin P. Bowen; Trent R. Northen; Dominique Loqué
• Keywords: 0301 basic medicine; 570; [SDV]Life Sciences [q-bio]; Science; Genetic Vectors; Bioengineering; Genetically Modified; Saccharomyces cerevisiae; Plant Roots; 630; Article; Industrial Biotechnology; 03 medical and health sciences; Genetics; Homologous Recombination; 2. Zero hunger; Q; Reproducibility of Results; Biological Sciences; Plants; Plants, Genetically Modified; Plant Leaves; [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology; Synthetic Biology; [SDV.EE.IEO]Life Sciences [q-bio]/Ecology, environment/Symbiosis; Biotechnology

AbstractThe advent and growth of synthetic biology has demonstrated its potential as a promising avenue of research to address many societal needs. However, plant synthetic biology efforts have been hampered by a dearth of DNA part libraries, versatile transformation vectors and efficient assembly strategies. Here, we describe a versatile system (named jStack) utilizing yeast homologous recombination to efficiently assemble DNA into plant transformation vectors. We demonstrate how this method can facilitate pathway engineering of molecules of pharmaceutical interest, production of potential biofuels and shuffling of disease-resistance traits between crop species. Our approach provides a powerful alternative to conventional strategies for stacking genes and traits to address many impending environmental and agricultural challenges.