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YLC-assembly: large DNA assembly via yeast life cycle.
He, Bo; Ma, Yuan; Tian, Fangfang; Zhao, Guang-Rong; Wu, Yi; Yuan, Ying-Jin.
Afiliação
  • He B; Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
  • Ma Y; Frontiers Research Institute for Synthetic Biology, Tianjin University, Tianjin 300072, China.
  • Tian F; Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
  • Zhao GR; Frontiers Research Institute for Synthetic Biology, Tianjin University, Tianjin 300072, China.
  • Wu Y; Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
  • Yuan YJ; Frontiers Research Institute for Synthetic Biology, Tianjin University, Tianjin 300072, China.
Nucleic Acids Res ; 51(15): 8283-8292, 2023 08 25.
Article em En | MEDLINE | ID: mdl-37486765
As an enabling technique of synthetic biology, the scale of DNA assembly largely determines the scale of genetic manipulation. However, large DNA assembly technologies are generally cumbersome and inefficient. Here, we developed a YLC (yeast life cycle)-assembly method that enables in vivo iterative assembly of large DNA by nesting cell-cell transfer of assembled DNA in the cycle of yeast mating and sporulation. Using this method, we successfully assembled a hundred-kilobase (kb)-sized endogenous yeast DNA and a megabase (Mb)-sized exogenous DNA. For each round, over 104 positive colonies per 107 cells could be obtained, with an accuracy ranging from 67% to 100%. Compared with other Mb-sized DNA assembly methods, this method exhibits a higher success rate with an easy-to-operate workflow that avoid in vitro operations of large DNA. YLC-assembly lowers the technical difficulty of Mb-sized DNA assembly and could be a valuable tool for large-scale genome engineering and synthetic genomics.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Saccharomyces cerevisiae / Técnicas Genéticas / Biologia Sintética Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Saccharomyces cerevisiae / Técnicas Genéticas / Biologia Sintética Idioma: En Ano de publicação: 2023 Tipo de documento: Article