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. 1999 Jul-Aug;1(4):275-9.
doi: 10.1002/(SICI)1521-2254(199907/08)1:4<275::AID-JGM44>3.0.CO;2-M.

Intercellular spread of GFP-VP22

A Aints  1 , M S Dilber, C I Smith
Affiliations

Intercellular spread of GFP-VP22

A Aints et al. J Gene Med. 1999 Jul-Aug.

Abstract

Background: The herpes simplex virus type 1 (HSV-1) VP22 polypeptide has been reported to mediate intercellular trafficking of heterologous proteins fused to its C- or N-terminus, a feature making it a useful tool in bystander cell-targeted gene therapy.

Methods: Here we show, by detection of Green Fluorescent Protein (GFP) fused to VP22, its subcellular distribution in living producer (transfected) and recipient (non-transfected) cells as well as in cells after fixation. Four cell lines from different species were used.

Results: Different fractions of translocated GFP-VP22 fusion protein could be detected in fixed recipient cells by two different methods of fixation. Functional GFP in live recipient cells could not be detected.

Conclusions: Our study indicates that the VP22-chimeric protein transfer in its present form is suboptimal in terms of protein function. However, after fixation, the GFP signal in 100% of cells in a monolayer can be detected even at moderate transfection efficiency.

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