scholarly journals Combining Single RNA Sensitive Probes with Subdiffraction-Limited and Live-Cell Imaging Enables the Characterization of Virus Dynamics in Cells

ACS Nano ◽  
2013 ◽  
Vol 8 (1) ◽  
pp. 302-315 ◽  
Author(s):  
Eric Alonas ◽  
Aaron W. Lifland ◽  
Manasa Gudheti ◽  
Daryll Vanover ◽  
Jeenah Jung ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Virus Dynamics ◽  
In Cells

scholarly journals A versatile reporter system to monitor virus infected cells and its application to dengue virus and SARS-CoV-2

2020 ◽  
Author(s):  
Felix Pahmeier ◽  
Christoper J Neufeldt ◽  
Berati Cerikan ◽  
Vibhu Prasad ◽  
Costantin Pape ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Viral Replication ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Fluorescent Protein ◽  
Live Cell ◽  
Reporter System ◽  
Infected Cells ◽  
Positive Strand Rna

ABSTRACTPositive-strand RNA viruses have been the etiological agents in several major disease outbreaks over the last few decades. Examples of that are flaviviruses, such as dengue virus and Zika virus that cause millions of yearly infections and spread around the globe, and coronaviruses, such as SARS-CoV-2, which is the cause of the current pandemic. The severity of outbreaks caused by these viruses stresses the importance of virology research in determining mechanisms to limit virus spread and to curb disease severity. Such studies require molecular tools to decipher virus-host interactions and to develop effective interventions. Here, we describe the generation and characterization of a reporter system to visualize dengue virus and SARS-CoV-2 replication in live cells. The system is based on viral protease activity causing cleavage and nuclear translocation of an engineered fluorescent protein that is expressed in the infected cells. We show the suitability of the system for live cell imaging and visualization of single infected cells as well as for screening and testing of antiviral compounds. Given the modular building blocks, the system is easy to manipulate and can be adapted to any virus encoding a protease, thus offering a high degree of flexibility.IMPORTANCEReporter systems are useful tools for fast and quantitative visualization of viral replication and spread within a host cell population. Here we describe a reporter system that takes advantage of virus-encoded proteases that are expressed in infected cells to cleave an ER-anchored fluorescent protein fused to a nuclear localization sequence. Upon cleavage, the fluorescent protein translocates to the nucleus, allowing for rapid detection of the infected cells. Using this system, we demonstrate reliable reporting activity for two major human pathogens from the Flaviviridae and the Coronaviridae families: dengue virus and SARS-CoV-2. We apply this reporter system to live cell imaging and use it for proof-of-concept to validate antiviral activity of a nucleoside analogue. This reporter system is not only an invaluable tool for the characterization of viral replication, but also for the discovery and development of antivirals that are urgently needed to halt the spread of these viruses.

scholarly journals Characterization of a new class of blue-fluorescent lipid droplet markers for live-cell imaging in plants

2015 ◽  
Vol 34 (4) ◽  
pp. 655-665 ◽  
Author(s):  
Soujanya Kuntam ◽  
László G. Puskás ◽  
Ferhan Ayaydin
Keyword(s):  
Lipid Droplet ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
New Class ◽  
Fluorescent Lipid

scholarly journals Detection and Characterization of Protein Interactions In Vivo by a Simple Live-Cell Imaging Method

PLoS ONE ◽  
2013 ◽  
Vol 8 (5) ◽  
pp. e62195 ◽  
Author(s):  
Oriol Gallego ◽  
Tanja Specht ◽  
Thorsten Brach ◽  
Arun Kumar ◽  
Anne-Claude Gavin ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Imaging Method

Characterization of fluorescent synthetic epicocconone-based dye through advanced light microscopies for live cell imaging applications

2017 ◽  
Vol 141 ◽  
pp. 394-405 ◽  
Author(s):  
Damien Schapman ◽  
Caroline Perraudeau ◽  
Magalie Bénard ◽  
Thibault Gallavardin ◽  
Agathe Boulangé ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell

Flow cytometry and live cell imaging for characterization of cell surface markers on human tissue-derived progenitors

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S69-S70
Author(s):  
W.A. Bova ◽  
V.R. Mantripragada ◽  
V. Luangphakdy ◽  
G.F. Muschler
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Human Tissue ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Surface Markers ◽  
Cell Surface Markers

Fluorogenic Protein Probes with Red or Near-Infrared Emission for Genetically Targeted Live-Cell Imaging

2020 ◽  
Author(s):  
Sylvestre P. J. T. Bachollet ◽  
Cyril Addi ◽  
Jean-Maurice Mallet ◽  
Blaise Dumat
Keyword(s):  
Live Cell Imaging ◽  
Near Infrared ◽  
Cell Imaging ◽  
Infrared Emission ◽  
Live Cell ◽  
Molecular Rotor ◽  
Model Protein ◽  
Near Infrared Emission

A series of red-emitting and near-infrared fluorogenic protein probes based on push-pull molecular rotor structures was developed. After characterization of their optical properties using Bovine Serum Albumin as a model protein, they were conjugated to a halogenoalkane ligand in order to target the protein self-labeling tag HaloTag. The interaction with HaloTag was investigated in vitro and then the most promising probes were applied to live-cell imaging in wash-free conditions using fluorogenic and chemogenetic targeting of HaloTag fusion proteins.<br>

scholarly journals A Versatile Reporter System to Monitor Virus Infected Cells and Its Application to Dengue Virus and SARS-CoV-2

2020 ◽  
Author(s):  
Felix Pahmeier ◽  
Christopher J. Neufeldt ◽  
Berati Cerikan ◽  
Vibhu Prasad ◽  
Costantin Pape ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Fluorescent Protein ◽  
Live Cell ◽  
Nuclear Localization Sequence ◽  
Reporter System ◽  
Infected Cells ◽  
Positive Strand Rna

Positive-strand RNA viruses have been the etiological agents in several major disease outbreaks over the last few decades. Examples of this include flaviviruses, such as dengue virus and Zika virus that cause millions of yearly infections around the globe, and coronaviruses, such as SARS-CoV-2, the source of the current pandemic. The severity of outbreaks caused by these viruses stresses the importance of research aimed at determining methods to limit virus spread and to curb disease severity. Such studies require molecular tools to decipher virus-host interactions and to develop effective treatments. Here, we describe the generation and characterization of a reporter system that can be used to visualize and identify cells infected with dengue virus or SARS-CoV-2. This system is based on viral protease activity that mediates cleavage and nuclear translocation of an engineered fluorescent protein stably expressed in cells. We show the suitability of this system for live cell imaging, for visualization of single infected cells, and for screening and testing of antiviral compounds. With the integrated modular building blocks, this system is easy to manipulate and can be adapted to any virus encoding a protease, thus offering a high degree of flexibility. IMPORTANCE Reporter systems are useful tools for fast and quantitative visualization of virus infected cells within a host cell population. Here we describe a reporter system that takes advantage of virus-encoded proteases that are expressed in infected cells to cleave an ER-anchored fluorescent protein fused to a nuclear localization sequence. Upon cleavage, the GFP moiety translocates to the nucleus, allowing for rapid detection of the infected cells. Using this system, we demonstrate reliable reporting activity for two major human pathogens from the Flaviviridae and the Coronaviridae families: dengue virus and SARS-CoV-2. We apply this reporter system to live cell imaging and use it for proof-of-concept to validate antiviral activity of a nucleoside analogue. This reporter system is not only an invaluable tool for the characterization of viral replication, but also for the discovery and development of antivirals that are urgently needed to halt the spread of these viruses.

Generation and functional characterization of fluorescent, N-terminally tagged CB1 receptor chimeras for live-cell imaging

2007 ◽  
Vol 35 (2) ◽  
pp. 237-248 ◽  
Author(s):  
Neil A. McDonald ◽  
Christopher M. Henstridge ◽  
Christopher N. Connolly ◽  
Andrew J. Irving
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Functional Characterization ◽  
Cb1 Receptor ◽  
Live Cell ◽  
Receptor Chimeras

A small-sized benzothiazole–indolium fluorescent probe: the study of interaction specificity targeting c-MYC promoter G-quadruplex structures and live cell imaging

2020 ◽  
Vol 56 (95) ◽  
pp. 15016-15019
Author(s):  
Bo-Xin Zheng ◽  
Meng-Ting She ◽  
Wei Long ◽  
Yong-Yu Xu ◽  
Yi-Han Zhang ◽  
...  
Keyword(s):  
Anticancer Activity ◽  
Fluorescent Probe ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
G Quadruplex ◽  
In Cells

A small-sized and target-specific fluorescent probe reveals the presence of c-MYC DNA G4-structures in cells and shows anticancer activity.

scholarly journals Construction and Characterization of “Spinach” Array for mRNA Live Cell Imaging

2014 ◽  
Vol 106 (2) ◽  
pp. 494a
Author(s):  
Jichuan Zhang ◽  
Jingyi Fei ◽  
Benjamin J. Leslie ◽  
Kyu Young Han ◽  
Thomas E. Kuhlman ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell
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