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  • Title: A method of alphavirus replicon particle titration based on expression of functional replicase/transcriptase.
    Author: Gangolli SS, Vasilakis N, Kovacs GR, Zamb TJ, Kowalski J.
    Journal: J Virol Methods; 2003 May; 109(2):133-8. PubMed ID: 12711055.
    Abstract:
    Alphavirus replicon particles are being exploited for a variety of purposes both in vitro as gene expression vectors, and in vivo as vaccines or gene therapy vectors. There is a need for a simple and universal method of titration of replicon particles that is independent of expression of the foreign protein. We devised a method that uses modified vaccinia virus Ankara (MVA) as an indicator virus, to deliver a Venezuelan equine encephalitis virus (VEE) defective helper RNA encoding green fluorescent protein (GFP). Co-infection of cells with the MVA-based indicator and Venezuelan equine encephalitis virus replicon particles (VRP) results in expression of the GFP gene. VRP titer is readily determined by counting fluorescent cells.
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