Engineering the ribosomal DNA in a megabase synthetic chromosome

Weimin Zhang; Guanghou Zhao; Zhouqing Luo; Yicong Lin; Lihui Wang; Yakun Guo; Ann Wang; Shuangying Jiang; Qingwen Jiang; Jianhui Gong; Yun Wang; Sha Hou; Jing Huang; Tianyi Li; Yiran Qin; Junkai Dong; Qin Qin; Jiaying Zhang; Xinzhi Zou; Xi He; Li Zhao; Yibo Xiao; Meng Xu; Erchao Cheng; Ning Huang; Tong Zhou; Yue Shen; Roy Walker; Yisha Luo; Zheng Kuang; Leslie A. Mitchell; Kun Yang; Sarah M. Richardson; Yi Wu; Bing Zhi Li; Ying Jin Yuan; Huanming Yang; Jiwei Lin; Guo Qiang Chen; Qingyu Wu; Joel S. Bader; Yizhi Cai; Jef D. Boeke; Junbiao Dai

doi:10.1126/science.aaf3981

Engineering the ribosomal DNA in a megabase synthetic chromosome

Weimin Zhang, Guanghou Zhao, Zhouqing Luo, Yicong Lin, Lihui Wang, Yakun Guo, Ann Wang, Shuangying Jiang, Qingwen Jiang, Jianhui Gong, Yun Wang, Sha Hou, Jing Huang, Tianyi Li, Yiran Qin, Junkai Dong, Qin Qin, Jiaying Zhang, Xinzhi Zou, Xi HeLi Zhao, Yibo Xiao, Meng Xu, Erchao Cheng, Ning Huang, Tong Zhou, Yue Shen, Roy Walker, Yisha Luo, Zheng Kuang, Leslie A. Mitchell, Kun Yang, Sarah M. Richardson, Yi Wu, Bing Zhi Li, Ying Jin Yuan, Huanming Yang, Jiwei Lin, Guo Qiang Chen, Qingyu Wu, Joel S. Bader, Yizhi Cai, Jef D. Boeke, Junbiao Dai

Immunology

Research output: Contribution to journal › Article › peer-review

159 Scopus citations

Abstract

We designed and synthesized a 976,067-base pair linear chromosome, synXII, based on native chromosome XII in Saccharomyces cerevisiae. SynXII was assembled using a two-step method, specified by successive megachunk integration and meiotic recombination-mediated assembly, producing a functional chromosome in S. cerevisiae. Minor growth defect "bugs" detected in synXII, caused by deletion of tRNA genes, were rescued by introducing an ectopic copy of a single tRNA gene. The ribosomal gene cluster (rDNA) on synXII was left intact during the assembly process and subsequently replaced by a modified rDNA unit used to regenerate rDNA at three distinct chromosomal locations. The signature sequences within rDNA, which can be used to determine species identity, were swapped to generate a Saccharomyces synXII strain that would be identified as Saccharomyces bayanus by standard DNA barcoding procedures.

Original language	English (US)
Article number	eaaf3981
Journal	Science
Volume	355
Issue number	6329
DOIs	https://doi.org/10.1126/science.aaf3981
State	Published - Mar 10 2017

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Zhang, W, Zhao, G, Luo, Z, Lin, Y, Wang, L, Guo, Y, Wang, A, Jiang, S, Jiang, Q, Gong, J, Wang, Y, Hou, S, Huang, J, Li, T, Qin, Y, Dong, J, Qin, Q, Zhang, J, Zou, X, He, X, Zhao, L, Xiao, Y, Xu, M, Cheng, E, Huang, N, Zhou, T, Shen, Y, Walker, R, Luo, Y, Kuang, Z, Mitchell, LA, Yang, K, Richardson, SM, Wu, Y, Li, BZ, Yuan, YJ, Yang, H, Lin, J, Chen, GQ, Wu, Q, Bader, JS, Cai, Y, Boeke, JD & Dai, J 2017, 'Engineering the ribosomal DNA in a megabase synthetic chromosome', Science, vol. 355, no. 6329, eaaf3981. https://doi.org/10.1126/science.aaf3981

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abstract = "We designed and synthesized a 976,067-base pair linear chromosome, synXII, based on native chromosome XII in Saccharomyces cerevisiae. SynXII was assembled using a two-step method, specified by successive megachunk integration and meiotic recombination-mediated assembly, producing a functional chromosome in S. cerevisiae. Minor growth defect {"}bugs{"} detected in synXII, caused by deletion of tRNA genes, were rescued by introducing an ectopic copy of a single tRNA gene. The ribosomal gene cluster (rDNA) on synXII was left intact during the assembly process and subsequently replaced by a modified rDNA unit used to regenerate rDNA at three distinct chromosomal locations. The signature sequences within rDNA, which can be used to determine species identity, were swapped to generate a Saccharomyces synXII strain that would be identified as Saccharomyces bayanus by standard DNA barcoding procedures.",

author = "Weimin Zhang and Guanghou Zhao and Zhouqing Luo and Yicong Lin and Lihui Wang and Yakun Guo and Ann Wang and Shuangying Jiang and Qingwen Jiang and Jianhui Gong and Yun Wang and Sha Hou and Jing Huang and Tianyi Li and Yiran Qin and Junkai Dong and Qin Qin and Jiaying Zhang and Xinzhi Zou and Xi He and Li Zhao and Yibo Xiao and Meng Xu and Erchao Cheng and Ning Huang and Tong Zhou and Yue Shen and Roy Walker and Yisha Luo and Zheng Kuang and Mitchell, {Leslie A.} and Kun Yang and Richardson, {Sarah M.} and Yi Wu and Li, {Bing Zhi} and Yuan, {Ying Jin} and Huanming Yang and Jiwei Lin and Chen, {Guo Qiang} and Qingyu Wu and Bader, {Joel S.} and Yizhi Cai and Boeke, {Jef D.} and Junbiao Dai",

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AU - Zhao, Guanghou

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AU - Lin, Yicong

AU - Wang, Lihui

AU - Guo, Yakun

AU - Wang, Ann

AU - Jiang, Shuangying

AU - Jiang, Qingwen

AU - Gong, Jianhui

AU - Wang, Yun

AU - Hou, Sha

AU - Huang, Jing

AU - Li, Tianyi

AU - Qin, Yiran

AU - Dong, Junkai

AU - Qin, Qin

AU - Zhang, Jiaying

AU - Zou, Xinzhi

AU - He, Xi

AU - Zhao, Li

AU - Xiao, Yibo

AU - Xu, Meng

AU - Cheng, Erchao

AU - Huang, Ning

AU - Zhou, Tong

AU - Shen, Yue

AU - Walker, Roy

AU - Luo, Yisha

AU - Kuang, Zheng

AU - Mitchell, Leslie A.

AU - Yang, Kun

AU - Richardson, Sarah M.

AU - Wu, Yi

AU - Li, Bing Zhi

AU - Yuan, Ying Jin

AU - Yang, Huanming

AU - Lin, Jiwei

AU - Chen, Guo Qiang

AU - Wu, Qingyu

AU - Bader, Joel S.

AU - Cai, Yizhi

AU - Boeke, Jef D.

AU - Dai, Junbiao

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Engineering the ribosomal DNA in a megabase synthetic chromosome

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