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• W1973305672 abstract "A simple, quantitative assay for measuring the oncogenic potential of integrating vectors is needed in order to improve vector design and safety. In this study, we have developed a transient plasmid-based assay to measure the activation of a reporter gene by an adjacent vector provirus. Plasmid pACT contains a luciferase cassette driven by a minimal, enhancerless promoter, into which vector proviruses are inserted upstream for evaluation by luciferase assays and northern blots. In a comparison of analogous vectors based on murine leukemia virus (MLV), human immunodeficiency virus (HIV), and foamy virus (FV), we observed significant enhancer activity and read-through transcription from MLV proviruses, and significant read-through transcription from HIV proviruses. HIV and FV proviruses containing an internal MLV long-terminal repeat (LTR) promoter also had significant enhancer activity, which was not observed with an internal promoter from the murine phosphoglycerate kinase-1 gene, PGK. These results demonstrate that neighboring gene activation can be limited by using internal promoter(s) lacking enhancer activity, especially when present in an FV vector backbone that prevents read-through transcription. Although the pACT assay does not measure oncogenesis directly, it should be useful for screening vectors before more time-consuming and costly animal studies are undertaken." @default.
• W1973305672 created "2016-06-24" @default.
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• W1973305672 date "2008-03-01" @default.
• W1973305672 modified "2023-10-14" @default.
• W1973305672 title "A Rapid and Quantitative Assay for Measuring Neighboring Gene Activation by Vector Proviruses" @default.
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• W1973305672 doi "https://doi.org/10.1038/sj.mt.6300398" @default.
• W1973305672 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/18209733" @default.
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