High-titer recombinant adeno-associated virus production from replicating amplicons and herpes vectors deleted for glycoprotein H

Hum Gene Ther. 1999 Oct 10;10(15):2527-37. doi: 10.1089/10430349950016861.


Production of high-titer rAAV is essential for in vivo clinical application. One limiting factor may be the failure of existing systems to replicate the packaging genome in such a way that expression of Rep and Cap proteins is coordinately amplified. DISC-HSV (disabled single-cycle virus) is a genetically modified herpes simplex virus (HSV) that by deletion of glycoprotein H (gH) is infectious only if propagated in a complementing cell line. In this study, we have used DISC-HSV as a helper for rAAV replication, and have simulated to some extent the amplication of the rep and cap genomes seen in wtAAV infection by incorporating both these and vector sequences in HSV amplicons. Facilitated production of AAV Rep and Cap proteins translates into a considerably improved recovery of rAAV, which transduces cells of the neuroretina in vivo with high efficiency. The potential for contamination with infectious herpes particles is eliminated by the use of noncomplementing (gH-) cell lines to propagate the virus, and by standard purification methods. The use of DISC-HSV and herpes-derived amplicons for production of rAAV may be a useful strategy for future in vivo studies and for clinical application.

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • Cricetinae
  • Dependovirus / genetics*
  • Dependovirus / physiology
  • Gene Amplification*
  • Gene Deletion
  • Genetic Vectors*
  • Mice
  • Mice, Inbred BALB C
  • Plasmids
  • Recombination, Genetic
  • Simplexvirus / genetics*
  • Vero Cells
  • Viral Envelope Proteins / genetics*
  • Virus Replication


  • Viral Envelope Proteins
  • glycoprotein H, herpes simplex virus type 1