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.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.cell.2023.10.015 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Methodological advances enabled by the construction of a synthetic yeast genome.
Cell Rep Methods
April 2024
Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK. Electronic address:
The international Synthetic Yeast Project (Sc2.0) aims to construct the first synthetic designer eukaryote genome. Over the past few years, the Sc2.
View Article and Find Full Text PDFDesign, construction, and functional characterization of a tRNA neochromosome in yeast.
Cell
November 2023
Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK. Electronic address:
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.
View Article and Find Full Text PDFDebugging and consolidating multiple synthetic chromosomes reveals combinatorial genetic interactions.
Cell
November 2023
Institute for Systems Genetics and Department of Biochemistry and Molecular Pharmacology, NYU Langone Health, New York, NY 10016, USA; Department of Biomedical Engineering, NYU Tandon School of Engineering, Brooklyn, New York, NY 11201, USA. Electronic address:
The Sc2.0 project is building a eukaryotic synthetic genome from scratch. A major milestone has been achieved with all individual Sc2.
View Article and Find Full Text PDFThe Formation of Neochromosomes during Experimental Evolution in the Yeast .
Genes (Basel)
October 2021
Department of Genomes and Genetics, Institut Pasteur, CNRS UMR3525, F75724 Paris, France.
Novel, large-scale structural mutations were previously discovered during the cultivation of engineered strains in which essential tRNA synthetase genes were replaced by their orthologs from the distantly related yeast . Among those were internal segmental amplifications forming giant chromosomes as well as complex segmental rearrangements associated with massive amplifications at an unselected short locus. The formation of such novel structures, whose stability is high enough to propagate over multiple generations, involved short repeated sequences dispersed in the genome (as expected), but also novel junctions between unrelated sequences likely triggered by accidental template switching within replication forks.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!