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Total synthesis of a functional designer eukaryotic chromosome.

Narayana Annaluru 1 Héloïse Muller 2, 1, 3 Leslie A. Mitchell 4 Sivaprakash Ramalingam 5 Giovanni Stracquadanio 3, 6 Sarah M Richardson 6 Jessica S. Dymond 7 Zheng Kuang 3 Lisa Z. Scheifele 3, 8 Eric M. Cooper 3 Yizhi Cai 9, 10 Karen Zeller 3 Neta Agmon 3, 4 Jeffrey S. Han 11 Michalis Hadjithomas 12 Jennifer Tullman 6 Katrina Caravelli 3, 13 Kimberly Cirelli 1, 13 Zheyuan Guo 1, 14 Viktoriya London 1, 14 Apurva Yeluru 1, 14 Sindurathy Murugan 6 Karthikeyan Kandavelou 1, 15 Nicolas Agier 16 Gilles Fischer 16 Kun Yang 3, 6 J. Andrew Martin 3, 6 Murat Bilgel 14 Pavlo Bohutskyi 14 Kristin M. Boulier 13 Brian J. Capaldo 14 Joy Chang 14 Kristie Charoen 14 Woo Jin Choi 14 Peter Deng 12 James E Dicarlo 14 Judy Doong 14 Jessilyn Dunn 14 Jason I Feinberg 13 Christopher Fernandez 13 Charlotte E Floria 13 David Gladowski 13 Pasha Hadidi 14 Isabel Ishizuka 13 Javaneh Jabbari 13 Calvin Y. L. Lau 14 Pablo A. Lee 14 Sean Li 14 Denise Lin 13 Matthias E. Linder 13 Jonathan Ling 14 Jaime Liu 14 Jonathan Liu 14 Mariya London 13 Henry Ma 14 Jessica Mao 14 Jessica E Mcdade 14 Alex Mcmillan 13 Aaron M Moore 13 Won Chan Oh 14 Yu Ouyang 14 Ruchi Patel 14 Marina Paul 13 Laura C Paulsen 14 Judy Qiu 14 Alex Rhee 14 Matthew G Rubashkin 14 Ina Y. Soh 13 Nathaniel E Sotuyo 13 Venkatesh Srinivas 14 Allison Suarez 14 Andy Wong 14 Remus Wong 14 Wei Rose Xie 13 Yijie Xu 14 Allen T Yu 13 Romain Koszul 2 Joel S. Bader 3, 6 Jef D Boeke 3, 4, 12 Srinivasan Chandrasegaran 1
Abstract : 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 MATα allele on synIII. The complete design and synthesis of synIII establishes S. cerevisiae as the basis for designer eukaryotic genome biology.
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Submitted on : Tuesday, December 20, 2016 - 11:44:58 AM
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Narayana Annaluru, Héloïse Muller, Leslie A. Mitchell, Sivaprakash Ramalingam, Giovanni Stracquadanio, et al.. Total synthesis of a functional designer eukaryotic chromosome.. Science, American Association for the Advancement of Science, 2014, 344 (6179), pp.55-8. ⟨10.1126/science.1249252⟩. ⟨pasteur-01420002⟩

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