Generation of adenoviruses encoding the herpes simplex virus vhs gene: a novel strategy to generate adenoviruses expressing genes toxic to producer cells.
Adenovirus vectors expressing suicide genes represent a promising approach for cancer gene therapy. We wanted to determine whether the virion host shutoff (vhs) gene of herpes simplex virus could be used as a suicide gene for gene therapy of glioblastomas. The vhs gene was cloned downstream of the glial fibrillary acidic protein promoter to direct tissue-specific expression, and recombinant adenoviruses were generated in 293 cells. Viruses, which contained the vhs gene but did not express it, could readily be isolated; however, we were unable to plaque purify viruses that expressed vhs protein. We constructed a derivative of 293 cells expressing an antisense RNA to the vhs gene and used them to generate adenovirus vectors that express vhs protein. These cells were used to complement vhs-expressing adenoviruses through three rounds of plaque purification. This approach could be generalized to produce adenovirus vectors expressing other toxic transgenes.
Glenn
G M
GM
JCR Biopharmaceuticals, San Diego, California 92121, USA.
Chatterjee
S
S
D000256:Adenoviridae / Q000235:genetics* / Q000302:isolation & purification; D019380:Anti-HIV Agents / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D015316:Genetic Therapy; D005822:Genetic Vectors; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis / Q000235:genetics; D005909:Glioblastoma / Q000628:therapy*; D006561:Herpes Simplex / Q000201:enzymology*; D018259:Herpesvirus 1, Human / Q000201:enzymology*; D006801:Humans; D016133:Polymerase Chain Reaction; D016372:RNA, Antisense / Q000494:pharmacology; D011994:Recombinant Proteins / Q000009:adverse effects / Q000627:therapeutic use; D012260:Ribonucleases; D013937:Thymidine Kinase / Q000235:genetics*; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000378:metabolism
D019380:Anti-HIV Agents; D017931:DNA Primers; D005904:Glial Fibrillary Acidic Protein; D016372:RNA, Antisense; D011994:Recombinant Proteins; D014764:Viral Proteins; C070722:virion host shutoff protein, Simplexvirus; D013937:Thymidine Kinase; D012260:Ribonucleases