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Infrared laser–mediated gene induction in targeted single cells in vivo

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Abstract

We developed infrared laser–evoked gene operator (IR-LEGO), a microscope system optimized for heating cells without photochemical damage. Infrared irradiation causes reproducible temperature shifts of the in vitro microenvironment in a power-dependent manner. When applied to living Caenorhabditis elegans, IR-LEGO induced heat shock–mediated expression of transgenes in targeted single cells in a more efficient and less deleterious manner than a 440-nm dye laser and elicited physiologically relevant phenotypic responses.

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Figure 1: Schematic illustration of microscopic system for IR-LEGO, and thermal profiles of IR-laser irradiation in vitro.
Figure 2: IR laser–induced gene expression in vivo.

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Acknowledgements

We thank A. Fire (Stanford University), J. Hardin (University of Wisconsin), H. Kagoshima (National Institute of Genetics, Japan), Y. Kohara (National Institute of Genetics, Japan), K. Nishiwaki (Kwansei Gakuin University) and R. Tsien (University of California San Diego) for plasmids and W. Shoji (Tohoku Univeresity) for a transgenic zebrafish strain. We also thank Y. Shirasaki for helpful discussion, H. Omiya for technical advice about optics, A. Nukazuka for helping with early experiments on nematodes and Y. Oda for encouragement. mig-24(tk68) is a gift from K. Nishiwaki. This work was supported by the PRESTO and “Modelling of Innovative Research Results” research grants of Japan Science and Technology Agency, JST (S.Y.), the REIMEI Research Resources of Japan Atomic Energy Agency (Y.K.), The Sumitomo Foundation (S.T.), and grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (Y.K., T.T., S.T. and T.F.).

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Author notes

  1. Yasuhiro Kamei, Motoshi Suzuki and Kenjiro Watanabe: These authors contributed equally to this work.

Authors and Affiliations

  1. Radiation Biology Center, Kyoto University, Yoshida Konoe-cho, Sakyo-ku, 606-8501, Kyoto, Japan

    Yasuhiro Kamei & Takeshi Todo

  2. National Institute of Advanced and Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, 563-8577, Osaka, Japan

    Yasuhiro Kamei, Kazuhiro Fujimori, Takashi Kawasaki, Tomonori Deguchi & Shunsuke Yuba

  3. Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, 464-8602, Nagoya, Japan

    Motoshi Suzuki & Shin Takagi

  4. Major in Bioscience and Biomedical Engineering, Waseda University, 3-4-1 Okubo, Shinjuku, 169-8855, Tokyo, Japan

    Kenjiro Watanabe & Takashi Funatsu

  5. Department of Cell Biology and Neuroscience, Graduate School of Medicine, Osaka University, 2-2 Yamada-oka, Suita, 565-0871, Osaka, Japan

    Yoshihiro Yoneda

  6. Department of Radiation Biology and Medical Genetics, Graduate School of Medicine, Osaka University, 2-2 Yamada-oka, Suita, 565-0871, Osaka, Japan

    Takeshi Todo

  7. Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo, 113-0033, Tokyo, Japan

    Takashi Funatsu

Authors
  1. Yasuhiro Kamei
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  2. Motoshi Suzuki
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  3. Kenjiro Watanabe
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  4. Kazuhiro Fujimori
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  5. Takashi Kawasaki
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  6. Tomonori Deguchi
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  7. Yoshihiro Yoneda
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  8. Takeshi Todo
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  9. Shin Takagi
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  10. Takashi Funatsu
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  11. Shunsuke Yuba
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Corresponding authors

Correspondence to Yasuhiro Kamei or Shunsuke Yuba.

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Supplementary Text and Figures

Supplementary Figures 1–7, Supplementary Methods, Supplementary Discussion, Supplementary Note (PDF 1406 kb)

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Kamei, Y., Suzuki, M., Watanabe, K. et al. Infrared laser–mediated gene induction in targeted single cells in vivo. Nat Methods 6, 79–81 (2009). https://doi.org/10.1038/nmeth.1278

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