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

Kamei, Yasuhiro; Suzuki, Motoshi; Watanabe, Katsuyuki; Fujimori, Kazuhiro E.; Kawasaki, Takashi; Deguchi, Tomonori; Yoneda, Yoshihiro; Todo, Tomoki; Takagi, Shin; Funatsu, Takashi; Yuba, Shunsuke

doi:10.1038/nmeth.1278

Cited by 182 publications

(217 citation statements)

References 15 publications

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“…The IR laser-evoked gene operator (IR-LEGO) system is a new microscopic method optimized to heat cells by using an IR laser. We confirmed that this system was efficient in regulating spatiotemporal gene expression, and reported IR laser-mediated gene induction in single targeted cells in nematodes (Caenorhabditis elegans), 9 and the local gene expression in various tissues, such as the muscle, notochord, and retina in some living vertebrates, for example, zebrafish (Danio rerio) and medaka (Oryzias latipes). 10 In this study, we applied this IR-LEGO system to the vascular system in zebrafish and established, to the best of our knowledge, for the first time, an excellent method to induce laser-mediated gene expression in single targeted endothelial cells in vivo.…”

supporting

confidence: 63%

“…These movements seemed to influence the leakage of heat stress and made it difficult to diminish gene induction in nontargeted cells. Furthermore, applying the irradiating IR laser to the tissue resulted in intense heating of the elliptic region elongated vertically just before the focus, 9 suggesting that the muscle cells located in the vertical region of the optical axis were easily influenced by the laser. We attempted some modification of the IR-LEGO system by interrupting the center of the irradiated laser before focusing and tried to reduce the leakage to the surrounding cells in the vertical region.…”

Section: Discussionmentioning

confidence: 99%

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Application of Infrared Laser to the Zebrafish Vascular System

Kimura

Deguchi

Kamei

et al. 2013

ATVB

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Objective-Infrared laser-evoked gene operator is a new microscopic method optimized to heat cells in living organisms without causing photochemical damage. By combining the promoter system for the heat shock response, infrared laserevoked gene operator enables laser-mediated gene induction in targeted cells. We applied this method to the vascular system in zebrafish embryos and demonstrated its usability to investigate mechanisms of vascular morphogenesis in vivo. Approach and Results-We used double-transgenic zebrafish with fli1:nEGFP to identify the endothelial cells, and with hsp:mCherry to carry out single-cell labeling. Optimizing the irradiation conditions, we finally succeeded in inducing the expression of the mCherry gene in single targeted endothelial cells, at a maximum efficiency rate of 60%. In addition, we indicated that this system could be used for laser ablation under certain conditions. To evaluate infrared laser-evoked gene operator, we applied this system to the endothelial cells of the first intersegmental arteries, and captured images of the connection between the vascular systems of the brain and spinal cord. Conclusions-Our results suggest that the infrared laser-evoked gene operator system will contribute to the elucidation of the mechanisms underlying vascular morphogenesis by controlling spatiotemporal gene activation in

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supporting

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Application of Infrared Laser to the Zebrafish Vascular System

Kimura

Deguchi

Kamei

et al. 2013

ATVB

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“…In particular, optical regulation of gene expression (1)(2)(3)(4)(5)(6)(7)(8) and optogenetic control of receptor and channel proteins on the cell surface (9-13) have received much research attention. Heat shock proteins (HSPs) are critical components of a cell's defense mechanism against injury associated with adverse stresses (14)(15)(16)(17)(18).…”

mentioning

confidence: 99%

“…Heat shock proteins (HSPs) are critical components of a cell's defense mechanism against injury associated with adverse stresses (14)(15)(16)(17)(18). Promoters that contain heat shock elements (HSEs) upstream of the HSP coding region, known as heat shock promoters, can be turned on by various stresses besides heat shock, including heavy ions, radicals, alcohol, and viral infection, suggesting that they are driven by almost all kinds of stresses (1)(2)(3)(4)(5)(6)(7)(8)(14)(15)(16)(17)(18). Given the simple and highly controllable nature of heat shock treatment, the heat shock response is ubiquitous in all organisms from bacteria to higher vertebrates.…”

mentioning

confidence: 99%

Photothermic regulation of gene expression triggered by laser-induced carbon nanohorns

Miyako

Deguchi

Nakajima

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The development of optical methods to control cellular functions is important for various biological applications. In particular, heat shock promoter-mediated gene expression systems by laser light are attractive targets for controlling cellular functions. However, previous approaches have considerable technical limitations related to their use of UV, short-wavelength visible (vis), and infrared (IR) laser light, which have poor penetration into biological tissue. Biological tissue is relatively transparent to light inside the diagnostic window at wavelengths of 650–1,100 nm. Here we present a unique optical biotechnological method using carbon nanohorn (CNH) that transforms energy from diagnostic window laser light to heat to control the expression of various genes. We report that with this method, laser irradiation within the diagnostic window resulted in effective heat generation and thus caused heat shock promoter-mediated gene expression. This study provides an important step forward in the development of light-manipulated gene expression technologies.

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“…On one hand, the NIR laser heating, by water absorption to photons, actually stimulated the whole cells. For example, Y. Kamei et al showed quantitative analysis on thermal distribution by a NIR laser at 1480 nm in C. elegans and photothermal effect induced gene expression [75]. On the other hand, TRP channels were nonselective cation channels through which other cations could also enter cells.…”

Section: Cell Calcium Modulation By Optogenetics and Thermogeneticsmentioning

confidence: 99%

Regulation of Cellular Molecular Signaling by Light (Invited Paper)

Cheng¹,

Zhu²,

He³

2015

PIER

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Abstract-Laser technology has been promoting various microscopy methods and thus making great progresses in life science. Further than contribution to "seeing is believing", lasers have also demonstrated their capacity of manipulating cells and even molecular signaling. Specifically, with advances of lasers and combination with other techniques, recent reports show that cell calcium ion, a universal intra-and inter-cellular messenger, can be modulated by lasers at different levels of biological organization from organelle to tissue. It is very encouraging that laser irradiation can activate or control plenty of corresponding cell processes and functions by regulating cell calcium signaling pathways, with promising potential in both scientific research and clinical application. In this paper, optical techniques for regulation of cell calcium signaling are specifically reviewed. Most methods need exogenous chemicals or genetic materials to convert incident photon into stimulation that cells can response with specific molecular dynamics. The only all-optical approach is achieved by nonlinear excitation with femtosecond laser, despite lack of specificity and controllability, providing possibility of a totally noninvasive method without any biochemical materials and thus further potential clinical application in human beings. The developments and techniques of those methods are introduced and explained, with analysis on their properties and current challenges. Potential applications and prospective development are also discussed. Researchers on biophotonics and related biological fields can benefit from this review. It also provides a systematic reference to doctors and researchers who are working on practical application of those methods.

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

Cited by 182 publications

References 15 publications

Application of Infrared Laser to the Zebrafish Vascular System

Application of Infrared Laser to the Zebrafish Vascular System

Photothermic regulation of gene expression triggered by laser-induced carbon nanohorns

Regulation of Cellular Molecular Signaling by Light (Invited Paper)

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