Total Synthesis of a Functional Designer Eukaryotic Chromosome

被引：408

作者：

Annaluru, Narayana ^{[1
]}

Muller, Heloise ^{[1
,2
,3
,4
]}

Mitchell, Leslie A. ^{[2
,5
]}

Ramalingam, Sivaprakash ^{[1
]}

Stracquadanio, Giovanni ^{[2
,6
,7
]}

Richardson, Sarah M. ^{[6
,7
]}

Dymond, Jessica S. ^{[2
,8
]}

Kuang, Zheng ^{[2
]}

Scheifele, Lisa Z. ^{[2
,9
]}

Cooper, Eric M. ^{[2
]}

Cai, Yizhi ^{[2
,10
]}

Zeller, Karen ^{[2
]}

Agmon, Neta ^{[2
,5
]}

Han, Jeffrey S. ^{[11
]}

Hadjithomas, Michalis ^{[12
]}

Tullman, Jennifer ^{[6
,7
]}

Caravelli, Katrina ^{[2
,13
]}

Cirelli, Kimberly ^{[1
,13
]}

Guo, Zheyuan ^{[1
,14
]}

London, Viktoriya ^{[1
,14
]}

Yeluru, Apurva ^{[1
,14
]}

Murugan, Sindurathy ^{[6
,7
]}

Kandavelou, Karthikeyan ^{[1
,15
]}

Agier, Nicolas ^{[16
,17
]}

Fischer, Gilles ^{[16
,17
]}

Yang, Kun ^{[2
,6
,7
]}

Martin, J. Andrew ^{[2
,6
,7
]}

Bilgel, Murat ^{[14
]}

Bohutski, Pavlo ^{[14
]}

Boulier, Kristin M. ^{[13
]}

Capaldo, Brian J. ^{[14
]}

Chang, Joy ^{[14
]}

Charoen, Kristie ^{[14
]}

Choi, Woo Jin ^{[14
]}

Deng, Peter ^{[12
]}

DiCarlo, James E. ^{[14
]}

Doong, Judy ^{[14
]}

Dunn, Jessilyn ^{[14
]}

Feinberg, Jason I. ^{[13
]}

Fernandez, Christopher ^{[13
]}

Floria, Charlotte E. ^{[13
]}

Gladowski, David ^{[13
]}

Hadidi, Pasha ^{[14
]}

Ishizuka, Isabel ^{[13
]}

Jabbari, Javaneh ^{[13
]}

Lau, Calvin Y. L. ^{[14
]}

Lee, Pablo A. ^{[14
]}

Li, Sean ^{[14
]}

Lin, Denise ^{[13
]}

Linder, Matthias E. ^{[13
]}

机构：

[1] Johns Hopkins Univ, Sch Publ Hlth, Dept Environm Hlth Sci, Baltimore, MD 21205 USA

[2] Johns Hopkins Univ, Sch Med, High Throughput Biol Ctr, Baltimore, MD 21205 USA

[3] Inst Pasteur, Dept Genomes Genet, Grp Spatial Regulat Genomes, F-75015 Paris, France

[4] CNRS, UMR 3525, F-75015 Paris, France

[5] NYU, Langone Med Ctr, New York, NY 10016 USA

[6] Johns Hopkins Univ, Whiting Sch Engn, Dept Biomed Engn, Baltimore, MD 21218 USA

[7] Johns Hopkins Univ, Whiting Sch Engn, Inst Med Genet, Baltimore, MD 21218 USA

[8] Johns Hopkins Univ, Appl Phys Lab, Biol Sci Res & Exploratory Dev Dept, Laurel, MD 20723 USA

[9] Loyola Univ Maryland, Dept Biol, Baltimore, MD 21210 USA

[10] Univ Edinburgh, Edinburgh, Midlothian, Scotland

[11] Carnegie Inst Sci, Baltimore, MD 21218 USA

[12] Johns Hopkins Univ, Dept Biol, Baltimore, MD 21218 USA

[13] Johns Hopkins Univ, Krieger Sch Arts & Sci, Baltimore, MD 21218 USA

[14] Johns Hopkins Univ, Whiting Sch Engn, Baltimore, MD 21218 USA

[15] Pondicherry Biotech Private Ltd, Pillaichavady 605014, Puducherry, India

[16] Univ Paris 06, Sorbonne Univ, UMR 7238, F-75005 Paris, France

[17] CNRS, UMR 7238, F-75005 Paris, France

来源：

SCIENCE | 2014年 / 344卷 / 6179期

基金：

美国国家科学基金会;

关键词：

MYCOPLASMA-GENITALIUM GENOME; CHEMICAL-SYNTHESIS; YEAST; DNA; RECOMBINATION; PLASMID; CLONING; GENES; SITES;

D O I：

10.1126/science.1249252

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Rapid advances in DNA synthesis techniques have made it possible to engineer viruses, biochemical pathways and assemble bacterial genomes. Here, we report the synthesis of a functional 272,871-base pair designer eukaryotic chromosome, synIII, which is based on the 316,617-base pair native Saccharomyces cerevisiae chromosome III. Changes to synIII include TAG/TAA stop-codon replacements, deletion of subtelomeric regions, introns, transfer RNAs, transposons, and silent mating loci as well as insertion of loxPsym sites to enable genome scrambling. SynIII is functional in S. cerevisiae. Scrambling of the chromosome in a heterozygous diploid reveals a large increase in a-mater derivatives resulting from loss of the MAT alpha allele on synIII. The complete design and synthesis of synIII establishes S. cerevisiae as the basis for designer eukaryotic genome biology.

引用

页码：55 / 58

页数：4

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