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Establishing chromosomal design-build-test-learn through a synthetic chromosome and its combinatorial reconfiguration.pdf (6.63 MB)

Establishing chromosomal design-build-test-learn through a synthetic chromosome and its combinatorial reconfiguration

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posted on 2024-05-29, 04:38 authored by Jee Loon Foo, Shohei Kitano, Adelia Vicanatalita Susanto, Zhu Jin, Yicong Lin, Zhouqing Luo, Linsen Huang, Zhenzhen Liang, Leslie A. Mitchell, Kun Yang, Adison WongAdison Wong, Yizhi Cai, Jitong Cai, Giovanni Stracquadanio, Joel S. Bader, Jef D. Boeke, Junbiao Dai, Matthew Wook Chang

Chromosome-level design-build-test-learn cycles (chrDBTLs) allow systematic combinatorial reconfiguration of chromosomes with ease. Here, we established chrDBTL with a redesigned synthetic Saccharomyces cerevisiae chromosome XV, synXV. We designed and built synXV to harbor strategically inserted features, modified elements, and synonymously recoded genes throughout the chromosome. Based on the recoded chromosome, we developed a method to enable chrDBTL: CRISPR-Cas9-mediated mitotic recombination with endoreduplication (CRIMiRE). CRIMiRE allowed the creation of customized wild-type/synthetic combinations, accelerating genotype-phenotype mapping and synthetic chromosome redesign. We also leveraged synXV as a “build-to-learn” model organism for translation studies by ribosome profiling. We conducted a locus-to-locus comparison of ribosome occupancy between synXV and the wild-type chromosome, providing insight into the effects of codon changes and redesigned features on translation dynamics in vivo. Overall, we established synXV as a versatile reconfigurable system that advances chrDBTL for understanding biological mechanisms and engineering strains.

History

Journal/Conference/Book title

Cell Genomics

Publication date

2023-11-08

Version

  • Published

Corresponding author

Junbiao Dai; Matthew Wook Chang

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