Flavoprotein miniSOG as a genetically encoded photosensitizer for cancer cells

Ryumina, Alina P.; Serebrovskaya, Ekaterina O.; Shirmanova, Marina V.; Snopova, Ludmila B.; Кузнецова, М.; Turchin, Ilya V.; Ignatova, Nadezhda; Klementieva, Natalia V.; Fradkov, Arkady F.; Шахов, Б. Е.; Zagaynova, Elena V.; Lukyanov, Konstantin A.; Lukyanov, Sergey

doi:10.1016/j.bbagen.2013.07.015

Cited by 70 publications

(72 citation statements)

References 30 publications

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“…For live models, miniSOG and GFPlike protein KillerRed [13] (the latter produces radicalbased ROS under green-orange light) can be used as genetically encoded photosensitizers. It is a unique tool that enables very precise (in space and time) light-induced killing of specific cell populations [13][14][15][16][17], inactivation of target proteins [13,18 ,19], and damaging genomic DNA [16,20]. Moreover, a new approach to study protein arrangement in large complexes was developed [21 ].…”

Section: From Novel Fluorescent Proteins To Novel Applicationsmentioning

confidence: 99%

Novel uses of fluorescent proteins

Mishin

Belousov

Solntsev

et al. 2015

Current Opinion in Chemical Biology

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The field of genetically encoded fluorescent probes is developing rapidly. New chromophore structures were characterized in proteins of green fluorescent protein (GFP) family. A number of red fluorescent sensors, for example, for pH, Ca(2+) and H2O2, were engineered for multiparameter imaging. Progress in development of microscopy hardware and software together with specially designed FPs pushed superresolution fluorescence microscopy towards fast live-cell imaging. Deeper understanding of FPs structure and photophysics led to further development of imaging techniques. In addition to commonly used GFP-like proteins, unrelated types of FPs on the base of flavin-binding domains, bilirubin-binding domains or biliverdin-binding domains were designed. Their distinct biochemical and photophysical properties opened previously unexplored niches of FP uses such as labeling under anaerobic conditions, deep tissue imaging and even patients' blood analysis.

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Section: From Novel Fluorescent Proteins To Novel Applicationsmentioning

confidence: 99%

Novel uses of fluorescent proteins

Mishin

Belousov

Solntsev

et al. 2015

Current Opinion in Chemical Biology

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“…CALI has also been used as a 'switch' in optogenetics because it can disrupt chromatin and proteins in a locus-specific manner, kill specific organelles or a single cell in vivo Lin et al, 2013). Moreover, CALI is beginning to be exploited in photodynamic therapy of cancer cells (Mironova et al, 2013;Ryumina et al, 2013;Shirmanova et al, 2013;Liao et al, 2014). Therefore, we anticipate that the portfolio of applications of CALI will further expand, and contribute to a series of remarkable biological findings and medical applications in the future.…”

Section: Discussionmentioning

confidence: 99%

“…Fluorescence of miniSOG depends on the concentration of FMN; it is reduced when FMN levels decrease (Ryumina et al, 2013). FMN embedded in miniSOG protein enables the photoinduced electron-transfer reaction (type-I reaction), which results in a relatively poor yield of singlet oxygen compared with its free state, instead of generating singlet oxygen (type-II reaction) (see Box 1).…”

Section: Cali With Minisogmentioning

confidence: 99%

Chromophore-assisted laser inactivation – towards a spatiotemporal–functional analysis of proteins, and the ablation of chromatin, organelle and cell function

Sano

Watanabe

Matsunaga

2014

Journal of Cell Science

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Chromophore-assisted laser or light inactivation (CALI) has been employed as a promising technique to achieve spatiotemporal knockdown or loss-of-function of target molecules in situ. CALI is performed using photosensitizers as generators of reactive oxygen species (ROS). There are two CALI approaches that use either transgenic tags with chemical photosensitizers, or genetically encoded fluorescent protein fusions. Using spatially restricted microscopy illumination, CALI can address questions regarding, for example, protein isoforms, subcellular localization or phase-specific analyses of multifunctional proteins that other knockdown approaches, such as RNA interference or treatment with chemicals, cannot. Furthermore, rescue experiments can clarify the phenotypic capabilities of CALI after the depletion of endogenous targets. CALI can also provide information about individual events that are involved in the function of a target protein and highlight them in multifactorial events. Beyond functional analysis of proteins, CALI of nuclear proteins can be performed to induce cell cycle arrest, chromatin-or locus-specific DNA damage. Even at organelle levelsuch as in mitochondria, the plasma membrane or lysosomes -CALI can trigger cell death. Moreover, CALI has emerged as an optogenetic tool to switch off signaling pathways, including the optical depletion of individual neurons. In this Commentary, we review recent applications of CALI and discuss the utility and effective use of CALI to address open questions in cell biology.

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“…Stable CT26 cell lines were created using lentiviral transduction, essentially as described earlier (22). Briefly, the PCR fragment containing the mKate2-DEVD-iRFP or eqFP650-DEVD-iRFP open reading frame was cloned into Nde I- and Sal I-digested pRRLSIN.EF1.WPRE vector kindly provided by Didier Trono (Ecole Polytechnique Federale de Lausanne, Lausanne, Switzerland), with a modified multiple cloning site.…”

Section: Methodsmentioning

confidence: 99%

Genetically Encoded Far-Red Fluorescent Sensors for Caspase-3 Activity

et al. 2016

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Caspase-3 is a key effector caspase that is activated in both extrinsic and intrinsic pathways of apoptosis. Available fluorescent sensors for caspase-3 activity operate in relatively short wavelength regions and are nonoptimal for multiparameter microscopy and whole-body imaging. In the present work, we developed new genetically encoded sensors for caspase-3 activity possessing the most red-shifted spectra to date. These consist of Förster resonance energy transfer (FRET) pairs in which a far-red fluorescent protein (mKate2 or eqFP650) is connected to the infrared fluorescent protein iRFP through a linker containing the DEVD caspase-3 cleavage site. During staurosporine-induced apoptosis of mammalian cells (HeLa and CT26), both mKate2-DEVD-iRFP and eqFP650-DEVD-iRFP sensors showed a robust response (1.6-fold increase of the donor fluorescence intensity). However, eqFP650-DEVD-iRFP displayed aggregation in some cells. For stably transfected CT26 mKate2-DEVD-iRFP cells, fluorescence lifetime imaging (FLIM) enabled us to detect caspase-3 activation due to the increase of mKate2 donor fluorescence lifetime from 1.45 to 2.05 ns. We took advantage of the strongly red-shifted spectrum of mKate2-DEVD-iRFP to perform simultaneous imaging of EGFP-Bax translocation during apoptosis. We conclude that mKate2-DEVD-iRFP is well-suited for multiparameter imaging and also potentially beneficial for in vivo imaging in animal tissues.

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Flavoprotein miniSOG as a genetically encoded photosensitizer for cancer cells

Cited by 70 publications

References 30 publications

Novel uses of fluorescent proteins

Novel uses of fluorescent proteins

Chromophore-assisted laser inactivation – towards a spatiotemporal–functional analysis of proteins, and the ablation of chromatin, organelle and cell function

Genetically Encoded Far-Red Fluorescent Sensors for Caspase-3 Activity

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