Quantum Dot Labelling of Adenovirus Allows Highly Sensitive Single Cell Flow and Imaging Cytometry

Herod, Morgan R.; Pineda, Robert G.; Mautner, Vivien; Onion, David

doi:10.1002/smll.201401885

Cited by 12 publications

(9 citation statements)

References 76 publications

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“…This strategy is applicable to other Ad species or viral proteins . Alternatively, Ads have been labeled with biotin groups to couple quantum dots (Q‐dots), small particles containing highly photostable inorganic dyes via streptavidin . An ever‐expanding range of dyes with different excitation properties and ready‐to‐use kits has made direct capsid labeling a valuable and easy‐to‐use approach .…”

Section: General Considerations When Imaging Admentioning

confidence: 99%

Imaging the adenovirus infection cycle

Pied

Wodrich

2019

FEBS Letters

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Edited by Urs GreberIncoming adenoviruses seize control of cytosolic transport mechanisms to relocate their genome from the cell periphery to specialized sites in the nucleoplasm. The nucleus is the site for viral gene expression, genome replication, and the production of progeny for the next round of infection. By taking control of the cell, adenoviruses also suppress cell-autonomous immunity responses. To succeed in their production cycle, adenoviruses rely on wellcoordinated steps, facilitated by interactions between viral proteins and cellular factors. Interactions between virus and host can impose remarkable morphological changes in the infected cell. Imaging adenoviruses has tremendously influenced how we delineate individual steps in the viral life cycle, because it allowed the development of specific optical markers to label these morphological changes in space and time. As technology advances, innovative imaging techniques and novel tools for specimen labeling keep uncovering previously unseen facets of adenovirus biology emphasizing why imaging adenoviruses is as attractive today as it was in the past. This review will summarize past achievements and present developments in adenovirus imaging centered on fluorescence microscopy approaches.Adenoviruses (Ads) are nonenveloped icosahedral viruses with a diameter of~90 nm with slight structural differences between genotypes. The majority of the Ad capsid shell is composed of 240 hexon trimers, and each of the 12 vertices of the icosahedron is occupied by a penton. Pentons are involved in cell attachment and receptor recognition and are composed of the pentameric penton base from which the trimeric fiber molecule extends. Minor capsid proteins IIIa, VI, VIII, and IX are embedded in the capsid and contribute to capsid stability. Protein VI is located at the inner surface of the capsid, and biochemical data suggest that it may connect the capsid to the core containing the viral genome via protein V. The genome is ã 36-kb double-stranded linear DNA molecule with the terminal protein (TP) covalently bound to each 5 0end. Adenovirus genomes are highly condensed and organized into chromatin by several hundred copies of Abbreviations ADP, adenovirus death protein

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Section: General Considerations When Imaging Admentioning

confidence: 99%

Imaging the adenovirus infection cycle

Pied

Wodrich

2019

FEBS Letters

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“…Q-dots per se cannot pass through the lipid bilayer and have no inherent affinity to the major classes of cellular macromolecules, proteins, sugars, nucleic acids and lipids. Labeling viral particles with Q-dots through streptavidin-biotin interactions, functionalization with oligonucleotides or direct encapsidation in the virion have been utilized in studies with RSV, AdV, and HIV, and allowed virion tracking during entry into cells [ 107 , 108 , 109 , 110 , 111 ]. Other approaches included transfection, electroporation, and microinjection of Q-dots [ 112 ], for example to map HIV-1 provirus integration loci [ 113 ].…”

Section: Probes and Labeling Strategies For Imaging Of Virusesmentioning

confidence: 99%

Concepts in Light Microscopy of Viruses

Witte

Andriasyan

Georgi

et al. 2018

Viruses

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Viruses threaten humans, livestock, and plants, and are difficult to combat. Imaging of viruses by light microscopy is key to uncover the nature of known and emerging viruses in the quest for finding new ways to treat viral disease and deepening the understanding of virus–host interactions. Here, we provide an overview of recent technology for imaging cells and viruses by light microscopy, in particular fluorescence microscopy in static and live-cell modes. The review lays out guidelines for how novel fluorescent chemical probes and proteins can be used in light microscopy to illuminate cells, and how they can be used to study virus infections. We discuss advantages and opportunities of confocal and multi-photon microscopy, selective plane illumination microscopy, and super-resolution microscopy. We emphasize the prevalent concepts in image processing and data analyses, and provide an outlook into label-free digital holographic microscopy for virus research.

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“…Virus-labeling technology and single-virus tracking in real time are powerful tools for studying virus–host dynamics. These techniques allow visualization of successive events in the viral life cycle, which provides information that is missed during static imaging and are useful for elucidating the molecular mechanisms of virus–host interaction. − Compared with virus-labeling techniques such as fluorescent fusion protein expression and organic dye staining, inorganic semiconductor nanoparticle quantum dots (QDs) provide remarkable photostability, brightness, and biocompatibility for single-virus imaging and time-lapse tracking. , Many viruses, including human T-cell leukemia virus type 1, baculovirus, hematopoietic necrosis virus, influenza A virus, and adenovirus have been successfully labeled with QDs by direct virion decoration for imaging analyses. , However, previous reports of virus labeling with QDs have mainly modified peripheral viral components, such as the envelope or envelope glycoproteins, using chemical modification or direct biotinylation of the virion envelope.…”

Section: Preparation Of Hiv-1 Virions Encapsulating Qds By Site-speci...mentioning

confidence: 99%

Encapsulating Quantum Dots within HIV-1 Virions through Site-Specific Decoration of the Matrix Protein Enables Single Virus Tracking in Live Primary Macrophages

Yin

et al. 2018

Nano Lett.

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Labeling and imaging with quantum dots (QDs) provides powerful tools to visualize viral infection in living cells. Encapsulating QDs within virions represents a novel strategy for virus labeling. Here, we developed infectious HIV-1 virions encapsulating QDs through site-specific decoration of the viral matrix protein (MA) and used them to visualize early infection events in human primary macrophages by single-particle imaging. The MA protein was fused to a biotin acceptor peptide (BAP) tag, biotinylated, complexed with streptavidin-conjugated QDs in live cells, and incorporated into virions during virus assembly. The QD-encapsulated virions were tracked during infection of macrophages at a single particle level. The dynamic dissociation of MA and Vpr was also tracked in real time, and the results demonstrated that MA has multiple dynamic behaviors and functions during virus entry. More importantly, we tracked the dynamic interplay of QD-encapsulated virions with cellular mitochondria in live primary macrophages. We also found that HIV-1 can induce fission of mitochondria during the early phases of infection. In summary, we have constructed a type of QD-encapsulated virus particle and used this technology to further our understanding of the early events of HIV-1 infection.

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Quantum Dot Labelling of Adenovirus Allows Highly Sensitive Single Cell Flow and Imaging Cytometry

Cited by 12 publications

References 76 publications

Imaging the adenovirus infection cycle

Imaging the adenovirus infection cycle

Concepts in Light Microscopy of Viruses

Encapsulating Quantum Dots within HIV-1 Virions through Site-Specific Decoration of the Matrix Protein Enables Single Virus Tracking in Live Primary Macrophages

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