Total Synthesis of a Functional Designer Eukaryotic Chromosome

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Annaluru, N; Muller, H; Mitchell, LA; Ramalingam, S; Stracquadanio, G; Richardson, SM; Dymond, JS; Kuang, Z; Scheifele, LZ; Cooper, EM; Cai, Y; Zeller, K; Agmon, N; Han, JS; Hadjithomas, M; Tullman, J; Caravelli, K; Cirelli, K; Guo, Z; London, V; Yeluru, A; Murugan, S; Kandavelou, K; Agier, N; Fischer, G; Yang, K; Martin, JA; Bilgel, M; Bohutskyi, P; Boulier, KM; Capaldo, BJ; Chang, J; Charoen, K; Choi, WJ; Deng, P; DiCarlo, JE; Doong, J; Dunn, J; Feinberg, JI; Fernandez, C; Floria, CE; Gladowski, D; Hadidi, P; Ishizuka, I; Jabbari, J; Lau, CYL; Lee, PA; Li, S; Lin, D; Linder, ME; Ling, J; Liu, J; Liu, J; London, M; Ma, H; Mao, J; McDade, JE; McMillan, A; Moore, AM; Oh, WC; Ouyang, Y; Patel, R; Paul, M; Paulsen, LC; Qiu, J; Rhee, A; Rubashkin, MG; Soh, IY; Sotuyo, NE; Srinivas, V; Suarez, A; Wong, A; Wong, R; Xie, WR; Xu, Y; Yu, AT; Koszul, R; Bader, JS; Boeke, JD; Chandrasegaran, S

Published Date

  • April 4, 2014

Published In

Volume / Issue

  • 344 / 6179

Start / End Page

  • 55 - 58

Published By

Electronic International Standard Serial Number (EISSN)

  • 1095-9203

International Standard Serial Number (ISSN)

  • 0036-8075

Digital Object Identifier (DOI)

  • 10.1126/science.1249252

Language

  • en