title: A packaging cell line for propagation of recombinant adenovirus vectors containing two lethal gene-region deletions.
abstract: A cell line that provides the E1 as well as the E4 gene functions of human adenovirus 5 has been established by introduction of the full-length Ad5 E4 region into 293 cells. To avoid the E1A transactivation of the E4 gene expression, the E4 promoter was replaced by the mouse alpha inhibin promoter containing a cAMP response element. This cell line was used to generate E1/E4-deleted adenovirus vectors containing a lacZ gene in the E1 region under the control of mouse pgk promoter. The titer and the lacZ gene expression of E1/E4-deleted adenovirus vector were comparable to those of E1-deleted vectors. Evidence of cytopathic effect was absent following infection of nonpermissive cell lines with E1/E4-deleted adenovirus in vitro. Establishment of the 293-E4 cell line and the generation of E1/E4-deleted adenovirus vectors may prolong gene expression in vivo and significantly improve the safety of adenovirus vectors for human gene therapy.
authors: Cell Genesys Inc, Foster City, CA 94404, USA.
authors: Q
authors: Q
authors: Wang
authors: X C
authors: XC
authors: Jia
authors: M H
authors: MH
authors: Finer
mesh_terms: D017500:Adenovirus E1A Proteins / Q000096:biosynthesis / Q000235:genetics; D017504:Adenovirus E4 Proteins / Q000096:biosynthesis / Q000235:genetics; D000260:Adenoviruses, Human / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D046508:Culture Techniques / Q000379:methods; D017931:DNA Primers; D017353:Gene Deletion; D005804:Genes, Lethal; D005814:Genes, Viral; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D015533:Transcriptional Activation; D014162:Transfection
chemical_list: D017500:Adenovirus E1A Proteins; D017504:Adenovirus E4 Proteins; D017931:DNA Primers; D015345:Oligonucleotide Probes