Abstract
Epstein–Barr virus (EBV) evolved an episomal system for maintaining life-long, latent infection of human B lymphocytes. Circular episomes engineered from EBV components required for this latent form of infection have the capacity to persist in most types of replicating mammalian cells without DNA integration and the pitfalls of insertional mutagenesis. EBV episomes are typically transduced using low-efficiency methods. Here we present a method for efficient delivery of EBV episomes to nuclei of hepatocytes in living mice using a helper-dependent adenoviral vector and Cre-mediated recombination in vivo to generate circular EBV episomes following infection. Cre is transiently expressed from a hepatocyte-specific promoter so that vector generation and transgene expression are tissue specific. We show long-term persistence of the circularized vector DNA and expression of a reporter gene in hepatocytes of immunocompetent mice.
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Acknowledgements
This work was supported by USPHS Grant R37 CA025235 and the Eli and Edythe Broad Center of JSG was partially supported by the Ruth L Kirchstein National Research Service Award GM007185, and SDG by UCLA Training Grant in Genetic Mechanisms T32-GM07104.
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Gil, J., Gallaher, S. & Berk, A. Delivery of an EBV episome by a self-circularizing helper-dependent adenovirus: long-term transgene expression in immunocompetent mice. Gene Ther 17, 1288–1293 (2010). https://doi.org/10.1038/gt.2010.75
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DOI: https://doi.org/10.1038/gt.2010.75
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