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Precise programming of multigene expression stoichiometry in mammalian cells by a modular and programmable transcriptional system

Author

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  • Chenrui Qin

    (Peking University
    Peking University)

  • Yanhui Xiang

    (Chinese Academy of Sciences)

  • Jie Liu

    (Chinese Academy of Sciences)

  • Ruilin Zhang

    (Peking University)

  • Ziming Liu

    (Chinese Academy of Sciences)

  • Tingting Li

    (Chinese Academy of Sciences)

  • Zhi Sun

    (University of Chinese Academy of Science)

  • Xiaoyi Ouyang

    (Peking University)

  • Yeqing Zong

    (Bluepha Co., Ltd)

  • Haoqian M. Zhang

    (Bluepha Co., Ltd)

  • Qi Ouyang

    (Peking University)

  • Long Qian

    (Peking University)

  • Chunbo Lou

    (Chinese Academy of Sciences
    University of Chinese Academy of Science)

Abstract

Context-dependency of mammalian transcriptional elements has hindered the quantitative investigation of multigene expression stoichiometry and its biological functions. Here, we describe a host- and local DNA context-independent transcription system to gradually fine-tune single and multiple gene expression with predictable stoichiometries. The mammalian transcription system is composed of a library of modular and programmable promoters from bacteriophage and its cognate RNA polymerase (RNAP) fused to a capping enzyme. The relative expression of single genes is quantitatively determined by the relative binding affinity of the RNAP to the promoters, while multigene expression stoichiometry is predicted by a simple biochemical model with resource competition. We use these programmable and modular promoters to predictably tune the expression of three components of an influenza A virus-like particle (VLP). Optimized stoichiometry leads to a 2-fold yield of intact VLP complexes. The host-independent orthogonal transcription system provides a platform for dose-dependent control of multiple protein expression which may be applied for advanced vaccine engineering, cell-fate programming and other therapeutic applications.

Suggested Citation

  • Chenrui Qin & Yanhui Xiang & Jie Liu & Ruilin Zhang & Ziming Liu & Tingting Li & Zhi Sun & Xiaoyi Ouyang & Yeqing Zong & Haoqian M. Zhang & Qi Ouyang & Long Qian & Chunbo Lou, 2023. "Precise programming of multigene expression stoichiometry in mammalian cells by a modular and programmable transcriptional system," Nature Communications, Nature, vol. 14(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-37244-y
    DOI: 10.1038/s41467-023-37244-y
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    References listed on IDEAS

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    1. Yeqing Zong & Haoqian M. Zhang & Cheng Lyu & Xiangyu Ji & Junran Hou & Xian Guo & Qi Ouyang & Chunbo Lou, 2017. "Insulated transcriptional elements enable precise design of genetic circuits," Nature Communications, Nature, vol. 8(1), pages 1-13, December.
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    1. Roberto Di Blasi & Mara Pisani & Fabiana Tedeschi & Masue M. Marbiah & Karen Polizzi & Simone Furini & Velia Siciliano & Francesca Ceroni, 2023. "Resource-aware construct design in mammalian cells," Nature Communications, Nature, vol. 14(1), pages 1-10, December.

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