“Perfect” designer chromosome V and behavior of a ring derivative

作者：Ze‐Xiong Xie, Bing‐Zhi Li, Leslie A. Mitchell, Yi Wu, Xin Qi, Zhu Jin, Bin Jia, Xia Wang, Bo‐Xuan Zeng, Hui-Min Liu, Xiaole Wu, Q. Feng, Wen-Zheng Zhang, Wei Liu, Ming‐Zhu Ding, Xia Li, Guang-Rong Zhao, Jianjun Qiao, Jing‐Sheng Cheng, Meng Zhao, Zheng Kuang, Xuya Wang, James A. Martin, Giovanni Stracquadanio, Kun Yang, Xue Bai, Juan Zhao, Menglong Hu, Qiuhui Lin, Wenqian Zhang, Ming-Hua Shen, Si Chen, Wan Su, En‐Xu Wang, Rui Guo, Fang Zhai, Xue-Jiao Guo, Hao-Xing Du, Jia-Qing Zhu, Tian-Qing Song, Junjun Dai, Feifei Li, Guozhen Jiang, Shi-Lei Han, Shi-Yang Liu, Zhichao Yu, Xiaona Yang, Ken Chen, Cheng Hu, Da-Shuai Li, Nan Jia, Yue Liu, Lin-Ting Wang, Su Wang, Xiao-Tong Wei, Meiqing Fu, Lan-Meng Qu, Siyu Xin, Ting Liu, Kairen Tian, Xuenan Li, Jinhua Zhang, Li-Xiang Song, Jingui Liu, Jia-Fei Lv, Hang Xu, Ran Tao, Yan Wang, Tingting Zhang, Yexuan Deng, Yiran Wang, Ting Li, Guang-Xin Ye, Xiao-Ran Xu, Zheng-Bao Xia, Wei Zhang, Shi-Lan Yang, Yilin Liu, Wenqi Ding, Zhen-Ning Liu, Jun-Qi Zhu, Ningzhi Liu, Roy Walker, Yisha Luo, Yun Wang, Yue Shen, Huanming Yang, Yizhi Cai, Pingsheng Ma, Chun‐Ting Zhang, Joel S. Bader, Jef D. Boeke, Ying‐Jin Yuan · 发表于：Science · 年份：2017 · DOI：10.1126/science.aaf4704 · 被引用次数：264 · 研究领域：Fungal and yeast genetics research、CRISPR and Genetic Engineering、RNA and protein synthesis mechanisms

INTRODUCTION The Saccharomyces cerevisiae 2.0 project (Sc2.0) aims to modify the yeast genome with a series of densely spaced designer changes. Both a synthetic yeast chromosome arm (synIXR) and the entirely synthetic chromosome (synIII) function with high fitness in yeast. For designer genome synthesis projects, precise engineering of the physical sequence to match the specified design is important for the systematic evaluation of underlying design principles. Yeast can maintain nuclear chromosomes as rings, occurring by chance at repeated sequences, although the cyclized format is unfavorable in meiosis given the possibility of dicentric chromosome formation from meiotic recombination. Here, we describe the de novo synthesis of synthetic yeast chromosome V (synV) in the “Build-A-Genome China” course, perfectly matching the designer sequence and bearing loxPsym sites, distinguishable watermarks, and all the other features of the synthetic genome. We generated a ring synV derivative with user-specified cyclization coordinates and characterized its performance in mitosis and meiosis. RATIONALE Systematic evaluation of underlying Sc2.0 design principles requires that the final assembled synthetic genome perfectly match the designed sequence. Given the size of yeast chromosomes, synthetic chromosome construction is performed iteratively, and new mutations and unpredictable events may occur during synthesis; even a very small number of unintentional nucleotide changes across the ...