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Programmable mammalian translational modulators by CRISPR-associated proteins

Author

Listed:
  • Shunsuke Kawasaki

    (Kyoto University)

  • Hiroki Ono

    (Kyoto University
    Kyoto University)

  • Moe Hirosawa

    (Kyoto University)

  • Takeru Kuwabara

    (Kyoto University)

  • Shunsuke Sumi

    (Kyoto University
    Kyoto University)

  • Suji Lee

    (Kyoto University
    Kyoto University)

  • Knut Woltjen

    (Kyoto University)

  • Hirohide Saito

    (Kyoto University
    Kyoto University)

Abstract

Translational modulation based on RNA-binding proteins can be used to construct artificial gene circuits, but RNA-binding proteins capable of regulating translation efficiently and orthogonally remain scarce. Here we report CARTRIDGE (Cas-Responsive Translational Regulation Integratable into Diverse Gene control) to repurpose Cas proteins as translational modulators in mammalian cells. We demonstrate that a set of Cas proteins efficiently and orthogonally repress or activate the translation of designed mRNAs that contain a Cas-binding RNA motif in the 5’-UTR. By linking multiple Cas-mediated translational modulators, we designed and built artificial circuits like logic gates, cascades, and half-subtractor circuits. Moreover, we show that various CRISPR-related technologies like anti-CRISPR and split-Cas9 platforms could be similarly repurposed to control translation. Coupling Cas-mediated translational and transcriptional regulation enhanced the complexity of synthetic circuits built by only introducing a few additional elements. Collectively, CARTRIDGE has enormous potential as a versatile molecular toolkit for mammalian synthetic biology.

Suggested Citation

  • Shunsuke Kawasaki & Hiroki Ono & Moe Hirosawa & Takeru Kuwabara & Shunsuke Sumi & Suji Lee & Knut Woltjen & Hirohide Saito, 2023. "Programmable mammalian translational modulators by CRISPR-associated proteins," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
  • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-37540-7
    DOI: 10.1038/s41467-023-37540-7
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    References listed on IDEAS

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    Cited by:

    1. Yuanli Gao & Lei Wang & Baojun Wang, 2023. "Customizing cellular signal processing by synthetic multi-level regulatory circuits," Nature Communications, Nature, vol. 14(1), pages 1-14, December.

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