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Updated: May 6, 2026

Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
Published on: May 26, 2013
Tracking viral genomes in host cells at single-molecule resolution
I-Hsuan Wang1, Maarit Suomalainen, Vardan Andriasyan
1Institute of Molecular Life Sciences, University of Zürich, CH-8057 Zurich, Switzerland; Molecular Life Sciences Graduate School, ETH and University of Zürich, CH-8057 Zurich, Switzerland.
Researchers developed a new method to track viral DNA (vDNA) in single cells. This technique visualizes viral DNA trafficking, revealing nuclear import as a key bottleneck for adenovirus infection.
Area of Science:
- Molecular Biology
- Virology
- Cell Biology
Background:
- Viral DNA trafficking significantly influences cellular physiology and disease pathogenesis.
- Existing methods for visualizing and quantifying viral DNA (vDNA) trafficking lack single-molecule resolution and accuracy.
Purpose of the Study:
- To develop a noninvasive protocol for accurate quantification of viral DNA trafficking at single-cell resolution.
- To visualize and analyze viral DNA movement within mammalian cells during infection.
Main Methods:
- Metabolic labeling of newly synthesized viral DNA (vDNA) using ethynyl-modified nucleosides.
- Visualization via superresolution microscopy combined with copper(I)-catalyzed azide-alkyne cycloaddition (click) reactions.
- Application to adenovirus, herpes virus, and vaccinia virus infections in mammalian cells.
Main Results:
- Successful visualization of infection at single vDNA resolution.
- Demonstration of a large pool of capsid-free viral DNA accumulating in the cytosol during adenovirus uncoating.
- Identification of nuclear import of incoming viral DNA as a significant bottleneck in adenovirus infection.
Conclusions:
- The developed method provides unprecedented resolution for studying DNA virus replication cycles.
- This technique enables localization of cellular and viral factors interacting with viral DNA.
- It offers new avenues for studying innate immune responses to cytoplasmic viral DNA.
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