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Science. 2017 Mar 10;355(6329). pii: eaaf4704. doi: 10.1126/science.aaf4704.

"Perfect" designer chromosome V and behavior of a ring derivative.

Xie ZX1,2, Li BZ1,2, Mitchell LA3, Wu Y1,2, Qi X1,2, Jin Z1,2, Jia B1,2, Wang X1,2, Zeng BX1,2, Liu HM1,2, Wu XL1,2, Feng Q1,2, Zhang WZ1,2, Liu W1,2, Ding MZ1,2, Li X1,2, Zhao GR1,2, Qiao JJ1,2, Cheng JS1,2, Zhao M1,2, Kuang Z3, Wang X3, Martin JA3, Stracquadanio G4,5, Yang K4, Bai X1,2, Zhao J1,2, Hu ML1,2, Lin QH1,2, Zhang WQ1,2, Shen MH1,2, Chen S1,2, Su W1,2, Wang EX1,2, Guo R1,2, Zhai F1,2, Guo XJ1,2, Du HX1,2, Zhu JQ1,2, Song TQ1,2, Dai JJ1,2, Li FF1,2, Jiang GZ1,2, Han SL1,2, Liu SY1,2, Yu ZC1,2, Yang XN1,2, Chen K1,2, Hu C1,2, Li DS1,2, Jia N1,2, Liu Y1,2, Wang LT1,2, Wang S1,2, Wei XT1,2, Fu MQ1,2, Qu LM1,2, Xin SY1,2, Liu T1,2, Tian KR1,2, Li XN1,2, Zhang JH1,2, Song LX1,2, Liu JG1,2, Lv JF1,2, Xu H1,2, Tao R1,2, Wang Y1,2, Zhang TT1,2, Deng YX1,2, Wang YR1,2, Li T1,2, Ye GX1,2, Xu XR1,2, Xia ZB1,2, Zhang W1,2, Yang SL1,2, Liu YL1,2, Ding WQ1,2, Liu ZN1,2, Zhu JQ1,2, Liu NZ1,2, Walker R6, Luo Y6, Wang Y7, Shen Y7, Yang H7,8, Cai Y6, Ma PS1, Zhang CT1, Bader JS4, Boeke JD3, Yuan YJ9,2.

Author information

1
Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
2
SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin 300072, PR China.
3
Institute for Systems Genetics and Department of Biochemistry and Molecular Pharmacology, Langone Medical Center, New York University, New York City, NY 10016, USA.
4
High Throughput Biology Center and Department of Biomedical Engineering, Johns Hopkins University, Baltimore 21205, MD, USA.
5
School of Computer Science and Electronic Engineering, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, England, UK.
6
School of Biological Sciences, University of Edinburgh, Edinburgh EH9 3BF, Scotland, UK.
7
BGI-Shenzhen, Shenzhen 518083, PR China.
8
James D. Watson Institute of Genome Sciences, Hangzhou 310058, PR China.
9
Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China. yjyuan@tju.edu.cn.

Abstract

Perfect matching of an assembled physical sequence to a specified designed sequence is crucial to verify design principles in genome synthesis. We designed and de novo synthesized 536,024-base pair chromosome synV in the "Build-A-Genome China" course. We corrected an initial isolate of synV to perfectly match the designed sequence using integrative cotransformation and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated editing in 22 steps; synV strains exhibit high fitness under a variety of culture conditions, compared with that of wild-type V strains. A ring synV derivative was constructed, which is fully functional in Saccharomyces cerevisiae under all conditions tested and exhibits lower spore viability during meiosis. Ring synV chromosome can extends Sc2.0 design principles and provides a model with which to study genomic rearrangement, ring chromosome evolution, and human ring chromosome disorders.

PMID:
28280151
DOI:
10.1126/science.aaf4704
[Indexed for MEDLINE]

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