A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of Saccharomyces cerevisiae
Year: 2023
Type: article
Abstract: High Resolution Image Download MS PowerPoint Slide Synthetic biology toolkits are one of the core foundations on which the field has been built, facilitating and accelerating efforts to reprogram cells and organisms for diverse biotechnological applications. The yeast Saccharomyces cerevisiae, an important model and industrial organism, has benefited from a wide range of toolkits. In particular, the MoClo Yeast Toolkit (YTK) enables the fast and straightforward construction of multigene plasmids from a library of highly characterized parts for programming new cellular behavior in a more predictable manner. While YTK has cultivated a strong parts ecosystem and excels in plasmid construction, it is limited in the extent and flexibility with which it can create new strains of yeast. Here, we describe a new and improved toolkit, the Multiplex Yeast Toolkit (MYT), that extends the capabilities of YTK and addresses strain engineering limitations. MYT provides a set of new integration vectors and selectable markers usable across common laboratory strains, as well as additional assembly cassettes to increase the number of transcriptional units in multigene constructs, CRISPR-Cas9 tools for highly efficient multiplexed vector integration, and three orthogonal and inducible promoter systems for conditional programming of gene expression. With these tools, we provide yeast synthetic biologists with a powerful platform to take their engineering ambitions to exciting new levels.
Source: ACS Synthetic Biology
Authors: William M. Shaw, Ahmad S. Khalil, Tom Ellis
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $5,000
Funders: National Science Foundation, U.S. Department of Defense, Schmidt Family Foundation, National Institutes of Health, Engineering and Physical Sciences Research Council, Biotechnology and Biological Sciences Research Council, National Institute of Biomedical Imaging and Bioengineering, Division of Computing and Communication Foundations, Division of Emerging Frontiers
Awards: SemiSynBio-II: Hybrid Bio-Electronic Microfluidic Memory Arrays for Large Scale Testing and Remote Deployment, G5041453029, Towards Genomes-to-Design: Building and Testing a Minimal Essential Chromosome, G575848002, Engineering Fellowships for Growth: Advanced synthetic biology measurement to enable programmable functional biomaterials, G6316977547, Genome Organisation for Optimising Synthetic Secondary Metabolism, EPIGENETICALLY PROGRAMMED BIOLOGICAL MATERIALS, Synthetic circuit technologies for precision control and signal processing in mammalian cells
OpenAlex ID: W4388410709