Red fluorescent redox-sensitive biosensor Grx1-roCherry

Shokhina, Arina G.; Kostyuk, Alexander I.; Ermakova, Yulia G.; Panova, Anastasiya S.; Staroverov, Dmitry B; Egorov, Evgeny S.; Баранов, Михаил С.; Belle, Gijsbert J. van; Katschinski, Dörthe M.; Belousov, Vsevolod V.; Bilan, Dmitry S.

doi:10.1016/j.redox.2018.101071

Cited by 28 publications

(18 citation statements)

References 55 publications

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“…Over the past two decades, several ROS-sensitive fluorescent probes for monitoring ROS production in living cells have been developed. These include dihydrochlorofluorescein-diacetate, dihydrorhodamine, dihydroethidium, and HyPer family probes [25][26][27][28][29][30][31][32][33][34][35]. The advantages of using fluorescent probes include high sensitivity and method simplicity that allow direct quantification and localization of the sites of ROS formation within cell compartments.…”

Section: Discussionmentioning

confidence: 99%

Detection of Vascular Reactive Oxygen Species in Experimental Atherosclerosis by High-Resolution Near-Infrared Fluorescence Imaging Using VCAM-1-Targeted Liposomes Entrapping a Fluorogenic Redox-Sensitive Probe

Manea

Vlad

Rebleanu

et al. 2021

Oxidative Medicine and Cellular Longevity

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Excessive production of reactive oxygen species (ROS) and the ensuing oxidative stress are instrumental in all phases of atherosclerosis. Despite the major achievements in understanding the regulatory pathways and molecular sources of ROS in the vasculature, the specific detection and quantification of ROS in experimental models of disease remain a challenge. We aimed to develop a reliable and straightforward imaging procedure to interrogate the ROS overproduction in the vasculature and in various organs/tissues in atherosclerosis. To this purpose, the cell-impermeant ROS Brite™ 700 (RB700) probe that produces bright near-infrared fluorescence upon ROS oxidation was encapsulated into VCAM-1-targeted, sterically stabilized liposomes (VLp). Cultured human endothelial cells (EC) and macrophages (Mac) were used for in vitro experiments. C57BL6/J and ApoE-/- mice were randomized to receive normal or high-fat, cholesterol-rich diet for 10 or 32 weeks. The mice received a retroorbital injection with fluorescent tagged VLp incorporating RB700 (VLp-RB700). After two hours, the specific signals of the oxidized RB700 and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-DSPE), inserted into liposome bilayers, were measured ex vivo in the mouse aorta and various organs by high-resolution fluorescent imaging. VLp-RB700 was efficiently taken up by cultured human EC and Mac, as confirmed by fluorescence microscopy and spectrofluorimetry. After systemic administration in atherosclerotic ApoE-/- mice, VLp-RB700 were efficiently concentrated at the sites of aortic lesions, as indicated by the augmented NBD fluorescence. Significant increases in oxidized RB700 signal were detected in the aorta and in the liver and kidney of atherosclerotic ApoE-/- mice. RB700 encapsulation into sterically stabilized VCAM-1-sensitive Lp could be a novel strategy for the qualitative and quantitative detection of ROS in the vasculature and various organs and tissues in animal models of disease. The accurate and precise detection of ROS in experimental models of disease could ease the translation of the results to human pathologies.

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Section: Discussionmentioning

confidence: 99%

Detection of Vascular Reactive Oxygen Species in Experimental Atherosclerosis by High-Resolution Near-Infrared Fluorescence Imaging Using VCAM-1-Targeted Liposomes Entrapping a Fluorogenic Redox-Sensitive Probe

Manea

Vlad

Rebleanu

et al. 2021

Oxidative Medicine and Cellular Longevity

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“…Full-length mRNA samples were all synthesized using an SP6 kit (Life Technologies) from plasmid products after endonuclease digestion. After preparation, microinjection of mRNA was conducted into the yolk sac site close to the single cell of embryos at the 1-cell stage (before 0.5 hpf) [31]. The concentrations of mRNA samples used for microinjection were approximately 40–120 ng/μl, depending on experimental conditions.…”

Section: Methodsmentioning

confidence: 99%

The NOTCH1-dependent HIF1α/VGLL4/IRF2BP2 oxygen sensing pathway triggers erythropoiesis terminal differentiation

et al. 2020

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Hypoxia is widely considered as a limiting factor in vertebrate embryonic development, which requires adequate oxygen delivery for efficient energy metabolism, while nowadays some researches have revealed that hypoxia can induce stem cells so as to improve embryonic development. Erythroid differentiation is the oxygen delivery method employed by vertebrates at the very early step of embryo development, however, the mechanism how erythroid progenitor cell was triggered into mature erythrocyte is still not clear. In this study, after detecting the upregulation of vgll4b in response to oxygen levels, we generated vgll4b mutant zebrafish using CRISPR/Cas9, and verified the resulting impaired heme and dysfunctional erythroid terminal differentiation phenotype. Neither the vgll4b-deficient nor the γ-secretase inhibitor IX (DAPT)-adapted zebrafish were able to mediate HIF1α-induced heme generation. In addition, we showed that vgll4b mutant zebrafish were associated with an impaired erythroid phenotype, induced by the downregulation of alas2, which could be rescued by irf2bp2 depletion. Further mechanistic studies revealed that zebrafish VGLL4 sequesters IRF2BP2, thereby inhibiting its repression of alas2 expression and heme biosynthesis. These processes occur primarily via the VGLL4 TDU1 and IRF2BP2 ring finger domains. Our study also indicates that VGLL4 is a key player in the mediation of NOTCH1-dependent HIF1α-regulated erythropoiesis and can be sensitively regulated by oxygen concentrations. On the other hand, VGLL4 is a pivotal regulator of heme biosynthesis and erythroid terminal differentiation, which collectively improve oxygen metabolism.

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“…Reporter protein localized in vacuole and apoplast should be pH stable, a condition refusing many widely used fluorescent proteins [ 145 ]. For this reason, red variants of redox sensors roGFP and HyPer were designed, namely Grx1-roCherry [ 146 ] and HyPerRed [ 147 ], respectively. Additionally, HyPer and HyPerRed have their redox-non-sensitive and pH-sensitive counterparts and can be used in parallel as a control of pH effect on measurements [ 147 ].…”

Section: Challenges Of Plant Biosensors’ Development and Usementioning

confidence: 99%

Biosensors: A Sneak Peek into Plant Cell’s Immunity

Levak

Lukan

Gruden

et al. 2021

Life

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Biosensors are indispensable tools to understand a plant’s immunity as its spatiotemporal dimension is key in withstanding complex plant immune signaling. The diversity of genetically encoded biosensors in plants is expanding, covering new analytes with ever higher sensitivity and robustness, but their assortment is limited in some respects, such as their use in following biotic stress response, employing more than one biosensor in the same chassis, and their implementation into crops. In this review, we focused on the available biosensors that encompass these aspects. We show that in vivo imaging of calcium and reactive oxygen species is satisfactorily covered with the available genetically encoded biosensors, while on the other hand they are still underrepresented when it comes to imaging of the main three hormonal players in the immune response: salicylic acid, ethylene and jasmonic acid. Following more than one analyte in the same chassis, upon one or more conditions, has so far been possible by using the most advanced genetically encoded biosensors in plants which allow the monitoring of calcium and the two main hormonal pathways involved in plant development, auxin and cytokinin. These kinds of biosensor are also the most evolved in crops. In the last section, we examine the challenges in the use of biosensors and demonstrate some strategies to overcome them.

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Red fluorescent redox-sensitive biosensor Grx1-roCherry

Cited by 28 publications

References 55 publications

Detection of Vascular Reactive Oxygen Species in Experimental Atherosclerosis by High-Resolution Near-Infrared Fluorescence Imaging Using VCAM-1-Targeted Liposomes Entrapping a Fluorogenic Redox-Sensitive Probe

Detection of Vascular Reactive Oxygen Species in Experimental Atherosclerosis by High-Resolution Near-Infrared Fluorescence Imaging Using VCAM-1-Targeted Liposomes Entrapping a Fluorogenic Redox-Sensitive Probe

The NOTCH1-dependent HIF1α/VGLL4/IRF2BP2 oxygen sensing pathway triggers erythropoiesis terminal differentiation

Biosensors: A Sneak Peek into Plant Cell’s Immunity

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