Manipulating the 3D Organization of the Largest Synthetic Yeast Chromosome

• DOI: 10.2139/ssrn.4138701
• Publication Date: 2022-06-18T04:40:02Z
• Authors: Weimin Zhang; Luciana Lazar-Ste...; Hitoyoshi Yamashita; Michael J. Shen; Leslie A. Mitchell; Hikaru Kurasawa; Max A.B. Haase; Xiaoji Sun; Qingwen Jiang; Stephanie L. Lauer; Laura H. McCulloch; Yu Zhao; David M. Ichikawa; Nicole Easo; S. Jiaming Lin; Viola Fanfani; Brendan R. Camellato; Yinan Zhu; Jitong Cai; Zhuwei Xu; Maya Sacasa; Ryan Accardo; Ju Young Ahn; Surekha Annadanam; Leighanne A. Bram...; Nicholas R. Bello; Lousanna Cai; Stephanie Cerritos; MacIntosh Cornwell; Anthony D'Amato; Maria Hacker; Kenneth Hersey; Emma Kennedy; Ardeshir Kianercy; Dohee Kim; Hong Seo Lim; Griffin McCutcheon; Kimiko McGirr; Nora Meaney; Lauren Meyer; Ally Moyer; Maisa Nimer; Carla Sabbatini; Lisa Scheifele; Lucas Shores; Cassandra Silvest...; Arden Snee; Antonio Spina; Anthony Staiti; Matt Stuver; Elli Tian; Danielle Whearty; Calvin Zhao; Tony Zheng; Vivian Zhou; Karen Zeller; Joel S. Bader; Giovanni Stracqua...; Samuel Deutsch; Yasunori Aizawa; Junbiao Dai; Jef D. Boeke
• Keywords: 0301 basic medicine; 03 medical and health sciences

Whether synthetic genomes can power life has attracted broad interest in the biology field, especially when are extensively modified with thousands of designer features. Here we report de novo synthesis largest eukaryotic chromosome thus far, synIV, a 1,454,621-bp Saccharomyces cerevisiae resulting from extensive genome streamlining and modification. During construction developed megachunk assembly method, combined hierarchical integration strategy. This strategy significantly increased accuracy flexibility facilitated debugging. Additionally, further manipulated three-dimensional structure synIV nucleus to explore spatial gene regulation within nuclear space. Surprisingly, found few expression changes, suggesting that positioning inside yeast nucleoplasm plays minor role regulation. Therefore, our manipulation shed light on higher-order architectural design genomes.