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Related Experiment Videos

Intercellular spread of GFP-VP22.

A Aints1, M S Dilber, C I Smith

  • 1Department of Medicine, Clinical Research Centre, Huddinge Hospital, Karolinska Institutet, Sweden.

The Journal of Gene Medicine
|March 30, 2000
PubMed
Summary

Herpes simplex virus type 1 VP22 protein facilitates cell-to-cell transfer of fused proteins. While functional transfer is limited, fixed cells show high detection rates, aiding gene therapy applications.

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Area of Science:

  • Molecular Biology
  • Virology
  • Gene Therapy

Background:

  • Herpes simplex virus type 1 (HSV-1) VP22 protein mediates intercellular protein trafficking.
  • This property makes VP22 a potential tool for bystander cell-targeted gene therapy.

Purpose of the Study:

  • To investigate the subcellular distribution and functionality of Green Fluorescent Protein (GFP) fused to VP22.
  • To assess the efficiency of VP22-mediated protein transfer in different cell types.

Main Methods:

  • Fusion of GFP to VP22.
  • Detection of GFP-VP22 in living and fixed producer and recipient cells.
  • Utilized four different cell lines from various species.
  • Employed two distinct fixation methods for protein detection.

Main Results:

  • Translocated GFP-VP22 fusion protein was detected in fixed recipient cells.
  • Functional GFP was not detected in live recipient cells.
  • Fixed cells showed 100% GFP signal detection in monolayers at moderate transfection efficiencies.

Conclusions:

  • Current VP22-chimeric protein transfer is suboptimal for protein function.
  • Fixation significantly enhances the detectability of VP22-mediated protein transfer.
  • VP22-mediated transfer shows promise for gene therapy applications, particularly for detection in fixed cell populations.

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