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Fluorescently tagged canine adenovirus via modification with protein IX–enhanced green fluorescent protein

¹ Division of Human Gene Therapy, Departments of Medicine, Pathology and Surgery, and the Gene Therapy Center, University of Alabama at Birmingham, 901 19th Street South, BMR2-502, Birmingham, AL 35294, USA
² Departments of Pathology, Cell Biology and Surgery, University of Alabama at Birmingham, Birmingham, AL 35294-2172, USA

Correspondence
David T. Curiel
curiel{at}uab.edu

Canine adenovirus type 2 (CAV2) has become an attractive vector for gene therapy because of its non-pathogenicity and the lack of pre-existing neutralizing antibodies against this virus in the human population. Additionally, this vector has been proposed as a conditionally replicative adenovirus agent under the control of an osteocalcin promoter for evaluation in a syngeneic, immunocompetent canine model with spontaneous osteosarcoma. In this study, a CAV2 vector labelled with the fluorescent capsid fusion protein IX–enhanced green fluorescent protein (pIX–EGFP) was developed. Expression of the fluorescent fusion-protein label in infected cells with proper nuclear localization, and incorporation into virions, could be detected. The labelled virions could be visualized by fluorescence microscopy; this was applicable to the tracking of CAV2 infection, as well as localizing the distribution of the vector in tissues. Expression of pIX–EGFP could be exploited to detect the replication and spread of CAV2. These results indicate that pIX can serve as a platform for incorporation of heterologous proteins in the context of a canine adenovirus xenotype. It is believed that capsid-labelled CAV2 has utility for vector-development studies and for monitoring CAV2-based oncolytic adenovirus replication.

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				G. Schoehn, M. El Bakkouri, C. M. S. Fabry, O. Billet, L. F. Estrozi, L. Le, D. T. Curiel, A. V. Kajava, R. W. H. Ruigrok, and E. J. Kremer Three-Dimensional Structure of Canine Adenovirus Serotype 2 Capsid J. Virol., April 1, 2008; 82(7): 3192 - 3203. [Abstract] [Full Text] [PDF]