A multiplex MoClo Toolkit for extensive and flexible engineering of Saccharomyces cerevisiae
Author(s)
Shaw, William M
Khalil, Ahmad S
Ellis, Tom
Type
Journal Article
Abstract
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.
Date Issued
2023-11-17
Date Acceptance
2023-09-11
Citation
ACS Synthetic Biology, 2023, 12 (11), pp.3393-3405
ISSN
2161-5063
Publisher
American Chemical Society
Start Page
3393
End Page
3405
Journal / Book Title
ACS Synthetic Biology
Volume
12
Issue
11
Copyright Statement
Copyright © 2023 The Authors. Published by American Chemical Society. This publication is licensed under
CC-BY 4.0.
CC-BY 4.0.
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/37930278
Subjects
Biotechnology
CRISPR-Cas Systems
Ecosystem
Plasmids
Saccharomyces cerevisiae
CRISPR-Cas9
modular cloning
multiplex
Saccharomyces cerevisiae
synthetic biology
toolkit
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2023-11-06