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A photoactivatable Cre–loxP recombination system for optogenetic genome engineering

Nature Chemical Biology volume 12, pages 1059–1064 (2016)Cite this article

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Abstract

Genome engineering techniques represented by the Cre–loxP recombination system have been used extensively for biomedical research. However, powerful and useful techniques for genome engineering that have high spatiotemporal precision remain elusive. Here we develop a highly efficient photoactivatable Cre recombinase (PA-Cre) to optogenetically control genome engineering in vivo. PA-Cre is based on the reassembly of split Cre fragments by light-inducible dimerization of the Magnet system. PA-Cre enables sharp induction (up to 320-fold) of DNA recombination and is efficiently activated even by low-intensity illumination (∼0.04 W m−2) or short periods of pulsed illumination (∼30 s). We demonstrate that PA-Cre allows for efficient DNA recombination in an internal organ of living mice through noninvasive external illumination using a LED light source. The present PA-Cre provides a powerful tool to greatly facilitate optogenetic genome engineering in vivo.

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Figure 1: Design of a photoactivatable Cre–loxP recombination system.
Figure 2: Characterization of PA-Cre.
Figure 3: DNA recombination kinetics of PA-Cre.
Figure 4: PA-Cre activation with shorter periods of illumination.
Figure 5: PA-Cre-induced DNA recombination in mice.

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Acknowledgements

We thank Y. Nihongaki for his technical assistance in experiments with mice. We also thank S.E. Park, J.R. Quejada and S. Su for proofreading the manuscript. This work was supported by Platform for Dynamic Approaches to Living System from the Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT) and Japan Agency for Medical Research and Development (AMED). This work was also supported by a grant from the Takeda Science Foundation to M.S. This work was also supported by grants from Japan Society for the Promotion of Science (JSPS) to M.S. and JSPS Postdoctoral Fellowships for Research Abroad to F.K.

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  1. Fuun Kawano and Risako Okazaki: These authors contributed equally to this work.

Authors and Affiliations

  1. Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, Japan.,

    Fuun Kawano, Risako Okazaki & Moritoshi Sato

  2. Department of Rehabilitation and Regenerative Medicine, Columbia University, New York, USA

    Fuun Kawano & Masayuki Yazawa

  3. Department of Pharmacology, Columbia University, New York, USA

    Masayuki Yazawa

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Contributions

F.K., R.O. and M.S. conceived the project and designed the experiments. F.K. and R.O. performed the experiments. F.K., R.O. and M.S. wrote the manuscript. F.K., M.Y. and M.S. edited the manuscript and reviewed the data.

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Correspondence to Moritoshi Sato.

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The authors declare no competing financial interests.

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Kawano, F., Okazaki, R., Yazawa, M. et al. A photoactivatable Cre–loxP recombination system for optogenetic genome engineering. Nat Chem Biol 12, 1059–1064 (2016). https://doi.org/10.1038/nchembio.2205

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