2014
DOI: 10.1126/science.1249252
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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 bp designer eukaryotic chromosome, synIII, which is based on the 316,617 bp native Saccharomyces cerevisiae chromosome III. Changes to synIII include TAG/TAA stop-codon replacements, deletion of subtelomeric regions, introns, tRNAs, transposons and silent mating loci as well as insertion of loxPsym sites to enable genome… Show more

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Cited by 482 publications
(409 citation statements)
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“…1,5 1,5 ), as well as a suite of important new tools for genome editing (Box 1). With these combined advances the field is now at a point where gene synthesis and DNA assembly can empower even undergraduate students to construct entire eukaryotic chromosomes 6 .…”
Section: Introductionmentioning
confidence: 99%
“…1,5 1,5 ), as well as a suite of important new tools for genome editing (Box 1). With these combined advances the field is now at a point where gene synthesis and DNA assembly can empower even undergraduate students to construct entire eukaryotic chromosomes 6 .…”
Section: Introductionmentioning
confidence: 99%
“…Impressively, a brand new functional bacterial genome and eukaryotic chromosome have been successfully assembled using rational design and chemical synthesis. 33,34 Through random assembly of a set of constitutive promoters, a silent spectinabilin pathway from Streptomyces orinoci and a cryptic polycyclic tetramate macrolactams biosynthetic gene cluster from Streptomyces griseus were respectively discovered and characterized, 35,36 which confirmed the powerful ability of DNA-assembly methods in the discovery of novel natural products. More recently, using both combinatorial transcriptional engineering and directed evolution strategies, a library of promoters with varying strengths were assembled with the xylose-utilizing pathway or the cellobiose-utilizing pathway functional structural genes.…”
Section: Rapid Assembly Tools Enable Rapid Evolution Of Genes and Genmentioning
confidence: 81%
“…Whereas ARSs are typically spaced by 30-40 kb in S. cerevisiae (32), insertion of the 35-kb sequence carrying the glycolytic genes resulted in a spacing between adjacent confirmed ARSs (ARS504.2 and ARS507, and ARS912 and ARS913) of 82 and 74 kb for chromosomes V and IX, respectively (33). In their design for a synthetic chromosome III, Annaluru et al kept a conservative approach by maintaining 12 of the 19 native ARSs, with a maximum spacing between ARSs of ∼50 kb (34). Although it has been proposed that 120-300 kb of chromosomal DNA could be replicated from a single replication origin (35), a quantitative evaluation of the impact on the physiology of S. cerevisiae of increasing spacing between ARSs would facilitate the design of large chromosomal constructs and synthetic chromosomes.…”
Section: Discussionmentioning
confidence: 99%