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

Schindler, Daniel; Walker, Roy S.K.; Jiang, Shuangying; Brooks, Aaron N.; Wang, Yun; Müller, Carolin A.; Cockram, Charlotte; Luo, Yisha; García, Alicia; Schraivogel, Daniel; Mozziconacci, Julien; Blount, Benjamin A.; Cai, Jitong; Ogunlana, Lois; Liu, Wei; Jönsson, Katarina; Abramczyk, Dariusz; Garcia-Ruiz, Eva; Turowski, Tomasz W.; Swidah, Reem; Ellis, Tom; Antequera, Francisco; Shen, Yue; Nieduszynski, Conrad A.; Koszul, Romain; Dai, Junbiao; Steinmetz, Lars M.; Boeke, Jef D.; Cai, Yizhi

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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

Jitong Cai

Lois Ogunlana

Wei Liu

Katarina Jönsson

Dariusz Abramczyk

Eva Garcia-Ruiz

Tomasz W. Turowski

Reem Swidah

Tom Ellis

Francisco Antequera

Yue Shen

Conrad A. Nieduszynski

Romain Koszul

Junbiao Dai

Lars M. Steinmetz

Jef D. Boeke

Yizhi Cai


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.


Schindler, D., Walker, R. S., Jiang, S., Brooks, A. N., Wang, Y., Müller, C. A., …Cai, Y. Design, Construction, and Functional Characterization of a tRNA Neochromosome in Yeast

Publicly Available Date Oct 14, 2022
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Additional Information This bioRxiv article is a preprint and has not been certified by peer review.


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