Near-infrared fluorescent proteins

Shcherbo, Dmitry; Shemiakina, Irina I.; Ryabova, A. V.; Luker, Kathryn E.; Schmidt, Bradley T.; Souslova, Ekaterina A.; Gorodnicheva, Tatiana; Strukova, Lydia A.; Shidlovskiy, Konstantin M; Britanova, Olga V.; Zaraisky, Andrey G.; Lukyanov, Konstantin A.; Loschenov, Victor B.; Chudakov, Dmitriy M.

doi:10.1038/nmeth.1501

Cited by 193 publications

(169 citation statements)

References 15 publications

(19 reference statements)

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“…Far-red or nearinfrared reporter gene labeling of cancer cells better suit such whole body imaging applications (31). Accordingly, we expect whole-body, time-domain imaging of NTR reporter gene expression to broaden the possibilities of noninvasive optical imaging and be a practical and useful addition to current fluorescence and bioluminescence methodologies.…”

Section: Discussionmentioning

confidence: 99%

Nitroreductase, a Near-Infrared Reporter Platform for In Vivo Time-Domain Optical Imaging of Metastatic Cancer

McCormack

Silden

West³

et al. 2013

Cancer Research

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The ability to visualize reporter gene expression in vivo has revolutionized all facets of biologic investigation and none more so than imaging applications in oncology. Near-infrared reporter gene imaging may facilitate more accurate evaluation of chemotherapeutic response in preclinical models of orthotopic and metastatic cancers. We report the development of a cell permeable, quenched squarine probe (CytoCy5S), which is reduced by Escherichia coli nitroreductase (NTR), resulting in a near-infrared fluorescent product. Time-domain molecular imaging of NTR/CytoCy5S reporter platform permitted noninvasive monitoring of disease progression in orthotopic xenografts of disseminated leukemia, lung, and metastatic breast cancer. This methodology facilitated therapeutic evaluation of NTR gene-directed enzymatic prodrug therapy with conventional metronidazole antibiotics. These studies show NTR/CytoCy5S as a near-infrared gene reporter system with broad preclinical and prospective clinical applications within imaging, and gene therapy, of cancer. Cancer Res; 73(4); 1276-86. Ó2012 AACR.

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Section: Discussionmentioning

confidence: 99%

Nitroreductase, a Near-Infrared Reporter Platform for In Vivo Time-Domain Optical Imaging of Metastatic Cancer

McCormack

Silden

West³

et al. 2013

Cancer Research

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“…Most dual imaging models have been dualluciferase systems, but these systems can be technically challenging due to the delivery requirements of coelenterazine, the substrate required for expression of most Renilla and Gaussia luciferase reporters [16][17][18] . Fluorescent reporters have allowed easy monitoring of many cell lines and constructs in vitro, but has had limited success for in vivo imaging due to the overlap between tissue and fur autofluorescence and the emission spectra of many commonly used fluorescent reporters including GFP, DsRed, and TdTomato 15,19 . This concern has encouraged the development of far-red fluorescent proteins, which allow for better tissue penetrance and higher specific signal compared to background 15,19 .…”

Section: Introductionmentioning

confidence: 99%

“…Fluorescent reporters have allowed easy monitoring of many cell lines and constructs in vitro, but has had limited success for in vivo imaging due to the overlap between tissue and fur autofluorescence and the emission spectra of many commonly used fluorescent reporters including GFP, DsRed, and TdTomato 15,19 . This concern has encouraged the development of far-red fluorescent proteins, which allow for better tissue penetrance and higher specific signal compared to background 15,19 . TurboFP650, the fluorescent protein shown in this system, is far-red shifted and overcomes many of the issues involved with imaging fluorescent proteins in living animals.…”

Section: Introductionmentioning

confidence: 99%

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Development, Expansion, and <em>In vivo</em> Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and Induced Pluripotent Stem Cells (iPSCs)

Bock

Knorr

Kaufman

2013

JoVE

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We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to high levels of NK cells after 4 weeks culture and can undergo further 2-log expansion with artificial antigen presenting cells. hESCand iPSC-derived NK cells developed in this system have a mature phenotype and function. The production of large numbers of genetically modifiable NK cells is applicable for both basic mechanistic as well as anti-tumor studies. Expression of firefly luciferase in hESC-derived NK cells allows a non-invasive approach to follow NK cell engraftment, distribution, and function. We also describe a dual-imaging scheme that allows separate monitoring of two different cell populations to more distinctly characterize their interactions in vivo. This method of derivation, expansion, and dual in vivo imaging provides a reliable approach for producing NK cells and their evaluation which is necessary to improve current NK cell adoptive therapies. Video LinkThe video component of this article can be found at

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“…They are widely used to determine the spatiotemporal dynamics of intracellular molecules, organelles, and whole cells as they can provide high resolution images with great sensitivity and without causing significant cytotoxicity (1). Fluorescence-labeled cells, in particular, can be visualized using fluorescence microscopy, flow cytometry, and whole-body imaging techniques (2,3). Fluorescent probes have therefore become an essential tool to study biological events in living cells, tissues and animals (4,5).…”

Section: Introductionmentioning

confidence: 99%

Application of POLARIC™ fluorophores in an in vivo tumor model

et al. 2013

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Abstract. Fluorescent and luminescent tools are commonly used to study the dynamics of cancer progression and metastases in real-time. Fluorophores have become essential tools to study biological events. However, few can sustain fluorescence long enough during long-term studies. In the present study, we focused on a series of new amphiphilic fluorophores known as POLARIC™, which emit strong fluorescence in lipid bilayers and can be readily modified using the Suzuki-Miyaura cross-coupling reaction. Appropriate chemical modifications of substituent groups can improve target-site specificity, reduce cytotoxicity and prolong emission. Therefore, in contrast to conventional fluorescent probes, these fluorophores show promise for long-term monitoring of biological processes. In the present study, we conducted long-term observations of tumor growth and metastasis using a POLARIC derivative as a novel fluorescent probe. For this purpose, we studied the metastatic melanoma cell line A375-SM, which proliferates at a high rate. We compared the characteristics of the POLARIC probe with the commercially available fluorescent dye PKH26 and fluorescent protein mRFP1. A375-SM cells were labeled with these fluorescent probes and orthotopically implanted into nude mice. The fluorescence emitted by POLARIC was detected more than five weeks after implantation without causing detectable harmful effects on tumor growth. By contrast, fluorescence of cells labeled with PKH26 could not be detected at this same time. Furthermore, POLARIC-, but not PKH26-labeled cells, were also detected in lung metastases. These results indicate that labeling cells with POLARIC fluorophores can significantly extend the time course of in vivo studies on tumor cell growth. IntroductionFluorescent and luminescent tools are commonly used to study the dynamics of cancer progression and metastases in real-time. They are widely used to determine the spatiotemporal dynamics of intracellular molecules, organelles, and whole cells as they can provide high resolution images with great sensitivity and without causing significant cytotoxicity (1). Fluorescence-labeled cells, in particular, can be visualized using fluorescence microscopy, flow cytometry, and whole-body imaging techniques (2,3). Fluorescent probes have therefore become an essential tool to study biological events in living cells, tissues and animals (4,5).An important example of the utility of these techniques is the generation of genetically engineered cell lines that stably express natural fluorescent proteins such as GFP (6,7) introduced using retroviral vectors.A series of new amphiphilic fluorophores known as POLARIC™ has recently been reported (8,9). This series of fluorophores is now commercially available. These fluorophores provide advantages over other fluorescent dyes, such as strong fluorescence in lipid bilayers, and possess molecular structures that are readily modified using the Suzuki-Miyaura cross-coupling reaction. Thus, appropriate chemical modifications of substituent groups can imp...

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Near-infrared fluorescent proteins

Cited by 193 publications

References 15 publications

Nitroreductase, a Near-Infrared Reporter Platform for In Vivo Time-Domain Optical Imaging of Metastatic Cancer

Nitroreductase, a Near-Infrared Reporter Platform for In Vivo Time-Domain Optical Imaging of Metastatic Cancer

Development, Expansion, and <em>In vivo</em> Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and Induced Pluripotent Stem Cells (iPSCs)

Application of POLARIC™ fluorophores in an in vivo tumor model

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