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A bacterial phytochrome-based optogenetic system controllable with near-infrared light

Nature Methods volume 13, pages 591–597 (2016)Cite this article

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Abstract

Light-mediated control of protein–protein interactions to regulate cellular pathways is an important application of optogenetics. Here, we report an optogenetic system based on the reversible light-induced binding between the bacterial phytochrome BphP1 and its natural partner PpsR2 from Rhodopseudomonas palustris bacteria. We extensively characterized the BphP1–PpsR2 interaction both in vitro and in mammalian cells and then used this interaction to translocate target proteins to specific cellular compartments, such as the plasma membrane and the nucleus. We showed light-inducible control of cell morphology that resulted in a substantial increase of the cell area. We demonstrated light-dependent gene expression with 40-fold contrast in cultured cells, 32-fold in subcutaneous mouse tissue, and 5.7-fold in deep tissues in mice. Characteristics of the BphP1–PpsR2 optogenetic system include its sensitivity to 740- to 780-nm near-infrared light, its ability to utilize an endogenous biliverdin chromophore in eukaryotes (including mammals), and its spectral compatibility with blue-light-driven optogenetic systems.

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Figure 1: Spectral properties of BphP1 and characterization of BphP1–PpsR2 interaction in vitro.
Figure 2: Light-induced relocalization of BphP1 to the plasma membrane.
Figure 3: Light induction of cellular cytoskeletal rearrangements.
Figure 4: Recruitment of BphP1 to the nucleus and light-induced transcription activation.
Figure 5: Light-induced transcription activation in mice.

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Acknowledgements

We thank E. Giraud (Institute for Research and Development, Marseille), M. Papiz (Liverpool University), T. Beatty (University of British Columbia), W. Weber (University of Freiburg), P. Hackett (University of Minnesota), Z. Izsvak (Max Delbrück Center for Molecular Medicine), Y. Yang (East China University of Science and Technology), and S. Masuda (Tokyo Institute of Technology) for plasmids and D. Shcherbakova, K. Chernov, and T. Redchuk for useful suggestions. This work was sponsored by National Institutes of Health grants GM073913, GM108579, and CA164468 to V.V.V.

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  1. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA

    Andrii A Kaberniuk, Anton A Shemetov & Vladislav V Verkhusha

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  1. Andrii A Kaberniuk
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  2. Anton A Shemetov
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Contributions

A.A.K. and A.A.S. characterized the proteins in vitro, in mammalian cell culture, and in vivo. V.V.V. planned and directed the project and, together with A.A.K. and A.A.S., designed the experiments, analyzed the data, and wrote the manuscript.

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Correspondence to Vladislav V Verkhusha.

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The authors declare no competing financial interests.

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Supplementary Figures 1–15, Supplementary Tables 1 and 2, and Supplementary Note (PDF 2453 kb)

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Kaberniuk, A., Shemetov, A. & Verkhusha, V. A bacterial phytochrome-based optogenetic system controllable with near-infrared light. Nat Methods 13, 591–597 (2016). https://doi.org/10.1038/nmeth.3864

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