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Rapid blue-light–mediated induction of protein interactions in living cells

Nature Methods volume 7, pages 973–975 (2010)Cite this article

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Abstract

Dimerizers allowing inducible control of protein-protein interactions are powerful tools for manipulating biological processes. Here we describe genetically encoded light-inducible protein-interaction modules based on Arabidopsis thaliana cryptochrome 2 and CIB1 that require no exogenous ligands and dimerize on blue-light exposure with subsecond time resolution and subcellular spatial resolution. We demonstrate the utility of this system by inducing protein translocation, transcription and Cre recombinase–mediated DNA recombination using light.

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Figure 1: Mapping of interacting domains of CRY2 and CIB1.
Figure 2: Light-triggered translocation of CRY2 in mammalian cells.
Figure 3: Light-induced activation of transcription and DNA recombination.

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Acknowledgements

We thank B. Arenkiel and E. Spana for comments and I. Davison for design of a custom LED light. This work was supported in part by grants from the US National Institutes of Health (R01 NS039402 and R01 MH064748) and the Howard Hughes Medical Institute (to M.D.E.), and US National Institutes of Health (R01 DK81584) (to C.L.T.).

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Author notes

  1. Matthew J Kennedy and Robert M Hughes: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neurobiology, Duke University Medical Center, Durham, North Carolina, USA

    Matthew J Kennedy, Joel W Schwartz & Michael D Ehlers

  2. Department of Biology, Duke University, Durham, North Carolina, USA

    Robert M Hughes, Leslie A Peteya & Chandra L Tucker

  3. Pfizer Global Research and Development, Neuroscience Research Unit, Groton, Connecticut, USA

    Michael D Ehlers

Authors
  1. Matthew J Kennedy
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  2. Robert M Hughes
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  4. Joel W Schwartz
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  6. Chandra L Tucker
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Contributions

C.L.T. conceived the idea and directed the work. M.J.K., R.M.H. and C.L.T. designed experiments. M.J.K., R.M.H., L.A.P., J.W.S. and C.L.T. performed experiments. M.D.E. supervised microscopy experiments. M.J.K., R.M.H. and C.L.T. wrote the manuscript. M.D.E. and C.L.T. edited the manuscript and reviewed the data.

Corresponding author

Correspondence to Chandra L Tucker.

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Competing interests

M.D.E. is an employee of Pfizer, Inc. A provisional patent application has been filed by Duke University on behalf of this technology.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 and Supplementary Table 1 (PDF 3395 kb)

Supplementary Table 2

Plasmids and oligos used in CRY2-CIB experiments (XLS 45 kb)

Supplementary Video 1

Light-triggered translocation of CRY2-mCh to the plasma membrane. HEK293T cells expressing CRY2-mCh were exposed to a 100-ms pulse of 488-nm light at t = 0. The mCherry channel was recorded at 3 Hz. The dimensions of the movie are 35 μm × 35 μm. (MOV 310 kb)

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Kennedy, M., Hughes, R., Peteya, L. et al. Rapid blue-light–mediated induction of protein interactions in living cells. Nat Methods 7, 973–975 (2010). https://doi.org/10.1038/nmeth.1524

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