A cell line for high-yield production of E1-deleted adenovirus vectors without the emergence of replication-competent virus.
Production of E1-deleted adenovirus vectors for gene therapy has been plagued by the emergence of replication-competent adenovirus. A number of investigators have minimized homologous sequences between the vector and transfected E1 DNA in an attempt to avoid replication-competent adenovirus. We describe a HeLa-based cell line called GH329 that stably expresses the E1 locus from a promoter derived from the phosphoglycerate kinase gene. Overlap sequences with a standard E1-deleted vector that retains a full pIX transcriptional unit have been eliminated at the 5' end and minimized at the 3' end. The GH329 cell line plaques and produces E1-deleted adenovirus as well as 293 cells. Replication-competent virus has emerged after 5 passages of vector on 293 cells but was not detected after 20 passages on GH329 cells.
Gao
G P
GP
Institute for Human Gene Therapy and Department of Molecular and Cellular Engineering, University of Pennsylvania, Philadelphia 19104, USA.
Engdahl
R K
RK
Wilson
J M
JM
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D002522:Chlorocebus aethiops; D003673:Defective Viruses / Q000235:genetics* / Q000502:physiology; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000096:biosynthesis*; D006367:HeLa Cells; D006801:Humans; D011995:Recombination, Genetic; D014709:Vero Cells; D010948:Viral Plaque Assay