Design, construction, and functional characterization of a tRNA neochromosome in yeast

• DOI: 10.1016/j.cell.2023.10.015
• Publication Date: 2023-11-08T15:26:38Z
• Authors: Daniel Schindler; Roy S.K. Walker; Shuangying Jiang; Aaron N. Brooks; Yun Wang; Carolin A. Müller; Charlotte Cockram; Yisha Luo; Alicia García; Daniel Schraivogel; Julien Mozziconacci; Noah Pena; Mahdi Assari; María del Carmen ...; Yu Zhao; Alba Ballerini; Benjamin A. Blount; Jitong Cai; Lois Ogunlana; Wei Liu; Katarina Jönsson; Dariusz Abramczyk; Eva Garcia-Ruiz; Tomasz W. Turowski; Reem Swidah; Tom Ellis; Tao Pan; Francisco Antequera; Yue Shen; Conrad A. Niedusz...; Romain Koszul; Junbiao Dai; Lars M. Steinmetz; Jef D. Boeke; Yizhi Cai
• Keywords: Proteomics; 570; Yeast 2.0; Ploidy changes; Chromosomes, Artificial, Yeast/genetics; Gene Expression Profiling; Synthetic genomics; Cellular burden; Saccharomyces cerevisiae; Genome reorganization; 530; Neochromosome; Saccharomyces cerevisiae/genetics; RNA, Transfer; Sc2.0; Synthetic Biology; Genome, Fungal; Rational chromosome design; tRNA; Chromosomes, Artificial, Yeast; RNA, Transfer/genetics; Synthetic biology

Here, we report the design, construction, and characterization of a tRNA neochromosome, designer chromosome that functions as an additional, de novo counterpart to native complement Saccharomyces cerevisiae. Intending address one central design principles Sc2.0 project, ∼190-kb neochromosome houses all 275 relocated nuclear genes. To maximize stability, incorporates orthogonal genetic elements from non-S. cerevisiae yeast species. Furthermore, presence 283 rox recombination sites enables SCRaMbLE system. Following construction in yeast, obtained evidence potent selective force, manifesting spontaneous doubling cell ploidy. sequencing, transcriptomics, proteomics, nucleosome mapping, replication profiling, FISH, Hi-C were undertaken investigate questions behavior function. Its demonstrates remarkable tractability model opens up opportunities directly test hypotheses surrounding these essential non-coding RNAs.