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Design, Construction, and Functional Characterization of a tRNA Neochromosome in Yeast

Schindler, D and Walker, RSK and Jiang, S and Brooks, AN and Wang, Y and Muller, CA and Cockram, C and Luo, Y and Garcia, A and Schraivogel, D and Mozziconacci, J and Blount, BA and Cai, J and Ogunlana, L and Liu, Wei and Jönsson, K and Abramczyk, D and Garcia-Ruiz, E and Turowski, TW and Swidah, R and Ellis, T and Antequera, F and Shen, Y and Nieduszynski, CA and Koszul, R and Dai, J and Steinmetz, LM and Boeke, JD and Cai, Y (2022) Design, Construction, and Functional Characterization of a tRNA Neochromosome in Yeast. biorXiv . (Submitted)


Official URL: https://www.biorxiv.org/content/10.1101/2022.10.03...


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 incorporated orthogonal genetic elements from non-S. cerevisiae yeast species. Furthermore, the presence of 283 rox recombination sites enable an orthogonal SCRaMbLE system capable of adjusting tRNA abundance. Following construction, we obtained evidence of a potent selective force once the neochromosome was introduced into yeast cells, 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 new opportunities to directly test hypotheses surrounding these essential non-coding RNAs.

Item Type:Article
Subjects:Q Science > QH Natural history > QH301 Biology
Divisions:Department of Genetics
ID Code:2257
Deposited By: dr Tomasz Turowski
Deposited On:03 Mar 2023 11:12
Last Modified:03 Mar 2023 11:12

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