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A SNAP-tagged derivative of HIV-1--a versatile tool to study virus-cell interactions

Manon Eckhardt1, Maria Anders, Walter Muranyi

  • 1Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany.

Plos One
|July 30, 2011
PubMed

Insights

Researchers developed HIV(SNAP), a modified human immunodeficiency virus (HIV), for advanced live-cell imaging. This tool enables detailed visualization of HIV-cell interactions using SNAP-tag technology for enhanced fluorescence microscopy.

Area of Science:

  • Virology
  • Microscopy
  • Molecular Biology

Background:

  • Fluorescent proteins (FPs) are used for live-cell imaging of HIV, but have limitations in properties and multi-color setups.
  • Developing novel tools is crucial for understanding HIV's dynamic life cycle and cell interactions.

Purpose of the Study:

  • To construct and characterize a novel human immunodeficiency virus (HIV) derivative, HIV(SNAP), utilizing the SNAP-tag system.
  • To evaluate HIV(SNAP) as a tool for advanced fluorescence microscopy and analysis of HIV-cell interactions.

Main Methods:

  • Genetically engineered HIV to incorporate the SNAP-tag into the Gag polyprotein.
  • Assessed viral particle assembly, release, maturation, infectivity, and replication capacity.
  • Utilized SNAP-reactive dyes and stochastic optical reconstruction microscopy (STORM) for visualization.

Main Results:

  • HIV(SNAP) assembly, release, and maturation were not significantly impaired.
  • Modified virions were infectious, though replication capacity was reduced.
  • SNAP-tag enabled specific staining of viral polyproteins and visualization of virions and budding sites.

Conclusions:

  • HIV(SNAP) is a versatile tool for live-cell imaging and sub-diffraction microscopy of HIV.
  • This system enhances the analysis of HIV-cell interactions and viral dynamics.
  • The SNAP-tag offers greater flexibility compared to traditional fluorescent proteins for HIV research.

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