Conditional repression of the E2 transcription unit in E1-E3-deleted adenovirus vectors is correlated with a strong reduction in viral DNA replication and late gene expression in vitro.
An E1-E3-deleted recombinant adenovirus vector expressing the hybrid protein TetR-KRAB has been produced. In this virus, AdTG9562, the E2 transcription is regulated by TetR-KRAB and tetO sequences inserted in cis. In the absence of tetracycline, a strong reduction in E2A gene expression, viral DNA replication, and late gene expression was observed in noncomplementing A549 cells, and a reduction in viral growth was seen in the E1-expressing 293 cells. In contrast, there was no repression in the presence of the regulator tetracycline. We propose that regulation by TetR-KRAB is a valuable tool with which to study the effects of viral gene expression in vitro.
Rittner
K
K
Gene Therapy Department, Transgène S.A., Strasbourg, France.
Schultz
H
H
Pavirani
A
A
Mehtali
M
M
D000256:Adenoviridae / Q000235:genetics*; D017498:Adenovirus E1 Proteins / Q000235:genetics*; D017502:Adenovirus E2 Proteins / Q000235:genetics*; D017503:Adenovirus E3 Proteins / Q000235:genetics*; D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis*; D004268:DNA-Binding Proteins / Q000235:genetics; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D012097:Repressor Proteins / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D015678:Viral Structural Proteins
D017498:Adenovirus E1 Proteins; D017502:Adenovirus E2 Proteins; D017503:Adenovirus E3 Proteins; D036022:Capsid Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D051741:Kruppel-Like Transcription Factors; D012097:Repressor Proteins; D014157:Transcription Factors; D015678:Viral Structural Proteins; C018315:hexon capsid protein, Adenovirus; C043322:penton protein, adenovirus; C034811:tetracycline resistance-encoding transposon repressor protein