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A polycistronic system for multiplexed and precalibrated expression of multigene pathways in fungi.
Yue, Qun; Meng, Jie; Qiu, Yue; Yin, Miaomiao; Zhang, Liwen; Zhou, Weiping; An, Zhiqiang; Liu, Zihe; Yuan, Qipeng; Sun, Wentao; Li, Chun; Zhao, Huimin; Molnár, István; Xu, Yuquan; Shi, Shuobo.
Afiliação
  • Yue Q; Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing, China.
  • Meng J; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
  • Qiu Y; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
  • Yin M; Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing, China.
  • Zhang L; Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing, China.
  • Zhou W; University of Chinese Academy of Sciences, Beijing, China.
  • An Z; Texas Therapeutics Institute, the Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, USA.
  • Liu Z; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
  • Yuan Q; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
  • Sun W; Key Lab for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing, China.
  • Li C; Key Lab for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing, China.
  • Zhao H; Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, USA.
  • Molnár I; VTT Technical Research Centre of Finland, Espoo, Finland. istvan.molnar@vtt.fi.
  • Xu Y; Biotechnology Research Institute, The Chinese Academy of Agricultural Sciences, Beijing, China. xuyuquan@caas.cn.
  • Shi S; Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China. shishuobo@mail.buct.edu.cn.
Nat Commun ; 14(1): 4267, 2023 07 17.
Article em En | MEDLINE | ID: mdl-37460548
Synthetic biology requires efficient systems that support the well-coordinated co-expression of multiple genes. Here, we discover a 9-bp nucleotide sequence that enables efficient polycistronic gene expression in yeasts and filamentous fungi. Coupling polycistronic expression to multiplexed, markerless, CRISPR/Cas9-based genome editing, we develop a strategy termed HACKing (Highly efficient and Accessible system by CracKing genes into the genome) for the assembly of multigene pathways. HACKing allows the expression level of each enzyme to be precalibrated by linking their translation to those of host proteins with predetermined abundances under the desired fermentation conditions. We validate HACKing by rapidly constructing highly efficient Saccharomyces cerevisiae cell factories that express 13 biosynthetic genes, and produce model endogenous (1,090.41 ± 80.92 mg L-1 squalene) or heterologous (1.04 ± 0.02 mg L-1 mogrol) terpenoid products. Thus, HACKing addresses the need of synthetic biology for predictability, simplicity, scalability, and speed upon fungal pathway engineering for valuable metabolites.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Sistemas CRISPR-Cas / Edição de Genes Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Sistemas CRISPR-Cas / Edição de Genes Idioma: En Ano de publicação: 2023 Tipo de documento: Article