Optogenetic control of intracellular signaling pathways

Zhang, Kai; Cui, Bianxiao

doi:10.1016/j.tibtech.2014.11.007

Cited by 212 publications

(192 citation statements)

References 81 publications

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“…By using a group of photoactivatable proteins, which undergo conformational changes and interact with each other upon excitation by light at specific wavelengths, optogenetics has extended its modality into interrogating complex intracellular signaling networks (Kennedy et al, 2010;Levskaya et al, 2009;Wu et al, 2009;Yazawa et al, 2009;Zhou et al, 2012). Shortly after the report of light-gated ion channel-based neuronal firing control (Boyden et al, 2005), optogenetics has been successfully used to control a variety of biological events in cell culture and single-cell organisms (reviewed by Kim and Lin, 2013;Muller et al, 2015;Schmidt and Cho, 2015;Tischer and Weiner, 2014;Toettcher et al, 2011;Tucker, 2012;Zhang and Cui, 2015;Zoltowski and Gardner, 2011). More recently, several studies have tested its application in multicellular organisms (Hallett et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Reversible optogenetic control of kinase activity during differentiation and embryonic development

et al. 2016

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A limited number of signaling pathways are repeatedly used to regulate a wide variety of processes during development and differentiation. The lack of tools to manipulate signaling pathways dynamically in space and time has been a major technical challenge for biologists. Optogenetic techniques, which utilize light to control protein functions in a reversible fashion, hold promise for modulating intracellular signaling networks with high spatial and temporal resolution. Applications of optogenetics in multicellular organisms, however, have not been widely reported. Here, we create an optimized bicistronic optogenetic system using Arabidopsis thaliana cryptochrome 2 (CRY2) protein and the N-terminal domain of cryptochrome-interacting basic-helix-loop-helix (CIBN). In a proofof-principle study, we develop an optogenetic Raf kinase that allows reversible light-controlled activation of the Raf/MEK/ERK signaling cascade. In PC12 cells, this system significantly improves lightinduced cell differentiation compared with co-transfection. When applied to Xenopus embryos, this system enables blue lightdependent reversible Raf activation at any desired developmental stage in specific cell lineages. Our system offers a powerful optogenetic tool suitable for manipulation of signaling pathways with high spatial and temporal resolution in a wide range of experimental settings.

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Section: Introductionmentioning

confidence: 99%

Reversible optogenetic control of kinase activity during differentiation and embryonic development

et al. 2016

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“…Currently, several lightinduced dimerization (LID) systems, in which photoactivatable proteins are dimerized by light in a reversible manner, are available. Among them, the cryptochrome 2 (CRY2) and cryptochrome-interacting basic helix-loop-helix1 (CIB1) pair has been widely used to control cell signaling through its lightinduced heterodimerization (11)(12)(13). CRY2 associates with its binding partner CIB1 upon exposure to blue light within a few seconds, and the complex is dissociated under a dark condition within several minutes (14).…”

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confidence: 99%

Efficient synthesis of phycocyanobilin in mammalian cells for optogenetic control of cell signaling

Uda

Goto

Oda

et al. 2017

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

114

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Optogenetics is a powerful tool to precisely manipulate cell signaling in space and time. For example, protein activity can be regulated by several light-induced dimerization (LID) systems. Among them, the phytochrome B (PhyB)–phytochrome-interacting factor (PIF) system is the only available LID system controlled by red and far-red lights. However, the PhyB–PIF system requires phycocyanobilin (PCB) or phytochromobilin as a chromophore, which must be artificially added to mammalian cells. Here, we report an expression vector that coexpresses HO1 and PcyA with Ferredoxin and Ferredoxin-NADP+ reductase for the efficient synthesis of PCB in the mitochondria of mammalian cells. An even higher intracellular PCB concentration was achieved by the depletion of biliverdin reductase A, which degrades PCB. The PCB synthesis and PhyB–PIF systems allowed us to optogenetically regulate intracellular signaling without any external supply of chromophores. Thus, we have provided a practical method for developing a fully genetically encoded PhyB–PIF system, which paves the way for its application to a living animal.

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“…Recently developed optogenetic actuators enable optical control over transmembrane voltage [12][13][14] as well as control over transcription [15], protein translocation [16], aggregation [17], dissociation [18], and binding and release [19,20]. By combining optogenetic actuators and reporters in the same cell, one can probe the input-output properties of individual cells with high throughput and information content.…”

Section: Introductionmentioning

confidence: 99%

Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation

Werley

Chien

Cohen

2017

Biomed. Opt. Express

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Abstract:The rapid increase in the number and quality of fluorescent reporters and optogenetic actuators has yielded a powerful set of tools for recording and controlling cellular state and function. To achieve the full benefit of these tools requires improved optical systems with high light collection efficiency, high spatial and temporal resolution, and patterned optical stimulation, in a wide field of view (FOV). Here we describe our 'Firefly' microscope, which achieves these goals in a Ø6 mm FOV. The Firefly optical system is optimized for simultaneous photostimulation and fluorescence imaging in cultured cells. All but one of the optical elements are commercially available, yet the microscope achieves 10-fold higher light collection efficiency at its design magnification than the comparable commercially available microscope using the same objective. The Firefly microscope enables all-optical electrophysiology ('Optopatch') in cultured neurons with a throughput and information content unmatched by other neuronal phenotyping systems. This capability opens possibilities in disease modeling and phenotypic drug screening. We also demonstrate applications of the system to voltage and calcium recordings in human induced pluripotent stem cell derived cardiomyocytes.

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Optogenetic control of intracellular signaling pathways

Cited by 212 publications

References 81 publications

Reversible optogenetic control of kinase activity during differentiation and embryonic development

Reversible optogenetic control of kinase activity during differentiation and embryonic development

Efficient synthesis of phycocyanobilin in mammalian cells for optogenetic control of cell signaling

Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation

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