Fluorescently Tagged Canine Adenovirus via Modification With Protein IX-enhanced Green Fluorescent Protein

J Gen Virol. 2005 Dec;86(Pt 12):3201-3208. doi: 10.1099/vir.0.80968-0.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenoviruses, Canine / genetics*
  • Adenoviruses, Canine / growth & development
  • Animals
  • Capsid Proteins / genetics*
  • Cell Line
  • Dogs
  • Genetic Vectors*
  • Green Fluorescent Proteins / genetics*
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Models, Animal
  • Recombinant Fusion Proteins*
  • Staining and Labeling / methods*

Substances

  • Capsid Proteins
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • hexon capsid protein, Adenovirus
  • Green Fluorescent Proteins