Identification of novel insertion sites in the Ad5 genome That utilize the Ad splicing machinery for therapeutic gene expression

摘要: Therapeutic transgene expression from oncolytic viruses represents one approach to increasing the effectiveness of these agents as cancer therapeutics. In case adenovirus (Ad), however, genomic packaging capacity is constrained. To address this, we explored whether a transposon-based system could identify sites in viral genome where endogenous Ad promoters drive via splicing and still maintain replication virus. Using GFP reporter gene an E3-deleted target, tested three signals. RACE analysis confirmed that GFP-expressing Ads occurs traced major late promoter (MLP). Replacement transposon by equivalent splice acceptor–luciferase cassette same orientation substitute transgenes are also expressed MLP. Interestingly, insertion opposite resulted that, some cases, originated within ITR region genome. summary, acceptor sequences can be used control this genomically economical arming Ads.