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On the cutting edge: protease-based methods for sensing and controlling cell biology

Nature Methods volume 17, pages 885–896 (2020)Cite this article

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Abstract

Sequence-specific proteases have proven to be versatile building blocks for tools that report or control cellular function. Reporting methods link protease activity to biochemical signals, whereas control methods rely on engineering proteases to respond to exogenous inputs such as light or chemicals. In turn, proteases have inherent control abilities, as their native functions are to release, activate or destroy proteins by cleavage, with the irreversibility of proteolysis allowing sustained downstream effects. As a result, protease-based synthetic circuits have been created for diverse uses such as reporting cellular signaling, tuning protein expression, controlling viral replication and detecting cancer states. Here, we comprehensively review the development and application of protease-based methods for reporting and controlling cellular function in eukaryotes.

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Fig. 1: Protease-based sensor designs.
Fig. 2: Protease-based signaling pathways.
Fig. 3: Protease-mediated drug control of proteins.
Fig. 4: Protease-mediated light control of proteins.

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Acknowledgements

We thank members of the Lin lab, M. Westberg and Y.S. Kim (all at Stanford University) for helpful discussions. This work was supported by a Stanford Discovery Innovation Award and NIH grant 1R21GM132687 to M.Z.L. H.K.C. is a Damon Runyon Fellow supported by Damon Runyon Cancer Research Foundation grant DRG-[2374-19].

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  1. H. Kay Chung

    Present address: The Salk Institute for Biological Studies, La Jolla, CA, USA

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  1. Department of Neurobiology, Stanford University, Stanford, CA, USA

    H. Kay Chung & Michael Z. Lin

  2. Department of Bioengineering, Stanford University, Stanford, CA, USA

    H. Kay Chung & Michael Z. Lin

  3. Program in Biological Sciences, Stanford University, Stanford, CA, USA

    H. Kay Chung

  4. Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA

    Michael Z. Lin

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Correspondence to Michael Z. Lin.

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H.K.C. is an inventor on patents describing SMASh and RASER. M.Z.L. is an inventor on patents describing SMASh, StaPL and RASER.

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Chung, H.K., Lin, M.Z. On the cutting edge: protease-based methods for sensing and controlling cell biology. Nat Methods 17, 885–896 (2020). https://doi.org/10.1038/s41592-020-0891-z

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