Total Synthesis of a Functional Designer Eukaryotic Chromosome

Journal Article (Academic 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 bp designer eukaryotic chromosome, synIII, which is based on the 316,617 bp native Saccharomyces cerevisiae chromosome III. Changes to synIII include TAG/TAA stopcodon replacements, deletion of subtelomeric regions, introns, tRNAs, 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 total synthesis of synIII represents the first complete design and synthesis of a eukaryotic chromosome, establishing 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; 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 2014

Published In

  • Science

Volume / Issue

  • 344 /

Chapter

  • 6179

Start / End Page

  • 55 - 58

Digital Object Identifier (DOI)

  • 10.1126/science.1249252

Language

  • en