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Related Concept Videos

Viral Mutations00:36

Viral Mutations

40.7K
A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material...
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Related Experiment Video

Updated: Mar 29, 2026

An Efficient Method for Adenovirus Production
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An Efficient Method for Adenovirus Production

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Directed Evolution of Adenoviruses.

Jason G Smith1

  • 1Department of Microbiology, University of Washington, Box 357735 1705 NE Pacific Street, Seattle, WA, 98195, USA. jgsmith2@uw.edu.

Methods in Molecular Biology (Clifton, N.J.)
|November 28, 2015
PubMed
Summary

Evolving viruses with reduced polymerase fidelity helps identify antiviral mechanisms. This study details methods to evolve a modified human adenovirus serotype 5 (HAdV-5) under antiviral pressure for drug discovery.

Area of Science:

  • Virology
  • Molecular Biology
  • Antiviral Research

Background:

  • Viral evolution in cell culture is crucial for understanding antiviral mechanisms and viral properties.
  • Mutations affecting viral polymerase fidelity can be leveraged for studying drug resistance and action.
  • A modified human adenovirus serotype 5 (HAdV-5) with reduced polymerase fidelity was previously developed.

Purpose of the Study:

  • To describe methods for evolving a HAdV-5 vector with a mutated polymerase (HAdV-5.polF421Y) under antiviral selective pressure.
  • To utilize this evolved virus to elucidate the mechanisms of action of antiviral compounds.

Main Methods:

  • Incorporation of a fidelity-reducing mutation into the pol gene of an HAdV-5 vector.
  • Culturing the modified virus (HAdV-5.polF421Y) under selective pressure from antiviral agents.
Keywords:
AdenovirusAntiviralEvolutionMutator

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  • Analyzing viral evolution to identify resistance mutations and infer drug mechanisms.

    • Main Results:

    • Successful evolution of HAdV-5.polF421Y under antiviral pressure.
    • Identification of specific viral adaptations that confer resistance to antivirals.
    • Insights into the molecular targets and mechanisms of action of tested antiviral compounds.

    Conclusions:

    • The described methods provide a powerful platform for discovering antiviral mechanisms of action.
    • Evolving viruses with reduced polymerase fidelity is a viable strategy for antiviral drug discovery and characterization.
    • This approach facilitates the elucidation of viral-drug interactions and resistance pathways.