Reconstruct a eukaryotic chromosome arm by de novo design and synthesis-猫眼课题宝

DOI：

10.1101/2022.10.04.509869

发表时间：

2022-10

期刊：

bioRxiv

影响因子：

通讯作者：

中科院分区：

其他

文献类型：

作者：

Shuangying Jiang;Zhouqing Luo;Kang Yu;Shijun Zhao;Zelin Cai;Wenfei Yu;Hong Wang;Li Cheng;Zhenzhen Liang;Hui Gao;M. Monti;Daniel Schindler;Linsen Huang;Cheng Zeng;Wei-Meng Zhang;Chun Zhou;Yuanwei Tang;Tianyi Li;Yingxin Ma;Yizhi Cai;J. Boeke;Junbiao Dai

生物体的基因组是从其祖先继承的，并随着时间的推移不断进化，然而，目前的版本可以改变多少仍然是未知的。在这里，我们使用染色体XII的左臂（chrXIIL）作为一个例子来探测酿酒酵母基因组的可塑性。新染色体被设计成容纳最初分散的基因。通过靶向DNA去除、染色体截短和随机缺失分析了chrXIIL中序列的重要性。值得注意的是，12个基因足以生存，而25个基因需要保持强大的适应性。接下来，我们证明了这些基因可以使用合成的调控序列和重新编码的开放阅读框与“一个氨基酸一个密码子”的策略进行重建。最后，我们构建了一个新的染色体，它可以取代chrXIIL的细胞活力，与这些重建的基因。我们的工作不仅突出了酵母基因组的高度可塑性，而且说明了用完全人工序列制造功能染色体的可能性。设计新染色体以促进外源DNA的组装以在酵母中稳定表达chrXII的左臂可以最小化到仅12个基因以维持活力，但需要额外的基因来保持强大的适应性使用人工调控序列的综合重新编码和转录重构产生功能性染色体臂完全重建的新染色体可以取代chrXIIL以保持可比性健身

The genome of an organism is inherited from its ancestor and keeps evolving over time, however, how much the current version could be altered remains unknown. Here, we use the left arm of chromosome XII (chrXIIL) as an example to probe the genome plasticity in Saccharomyces cerevisiae. A neochromosome was designed to harbor originally dispersed genes. The essentiality of sequences in chrXIIL was dissected by targeted DNA removal, chromosome truncation and random deletion. Notably, 12 genes were sufficient for survival, while 25 genes are required to retain robust fitness. Next, we demonstrated these genes could be reconstructed using synthetic regulatory sequences and recoded open-reading frames with “one-amino-acid-one-codon” strategy. Finally, we built a neochromsome, which could substitute for chrXIIL for cell viability, with these reconstructed genes. Our work not only highlights the high plasticity of yeast genome, but also illustrates the possibility of making functional chromosomes with completely artificial sequences. HIGHLIGHTS A neochromosome was designed to facilitate the assembly of exogenous DNA for stable expression in yeast The left arm of chrXII could be minimized to just 12 genes to maintain viability, but additional genes were required to retain robust fitness Comprehensive recoding and transcriptional refactoring using artificial regulatory sequences produced a functional chromosome arm A completely reconstructed neochromosome could replace the chrXIIL to maintain comparable fitness

Reconstruct a eukaryotic chromosome arm by de novo design and synthesis

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