Design, construction, and functional characterization of a tRNA neochromosome in yeast
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
Here, we report the design, construction, and characterization of a tRNA neochromosome, a designer chromosome that functions as an additional, de novo counterpart to the native complement of Saccharomyces cerevisiae. Intending to address one of the central design principles of the Sc2.0 project, the ∼190-kb tRNA neochromosome houses all 275 relocated nuclear tRNA genes. To maximize stability, the design incorporates orthogonal genetic elements from non-S. cerevisiae yeast species. Furthermore, the presence of 283 rox recombination sites enables an orthogonal tRNA SCRaMbLE system. Following construction in yeast, we obtained evidence of a potent selective force, manifesting as a spontaneous doubling in cell ploidy. Furthermore, tRNA sequencing, transcriptomics, proteomics, nucleosome mapping, replication profiling, FISH, and Hi-C were undertaken to investigate questions of tRNA neochromosome behavior and function. Its construction demonstrates the remarkable tractability of the yeast model and opens up opportunities to directly test hypotheses surrounding these essential non-coding RNAs.
Date Issued
2023-11-22
Date Acceptance
2023-10-12
Citation
Cell, 2023, 186 (24), pp.5237-5253.e22
ISSN
0092-8674
Publisher
Elsevier
Start Page
5237
End Page
5253.e22
Journal / Book Title
Cell
Volume
186
Issue
24
Copyright Statement
2023 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/37944512
PII: S0092-8674(23)01130-3
Subjects
Chromosomes, Artificial, Yeast
Gene Expression Profiling
Genome, Fungal
Proteomics
RNA, Transfer
Saccharomyces cerevisiae
Synthetic Biology
cellular burden
genome reorganization
neochromosome
ploidy changes
rational chromosome design
Sc2.0
synthetic biology
synthetic genomics
tRNA
Yeast 2.0
Publication Status
Published
Coverage Spatial
United States
Date Publish Online
2023-11-08