Effect of DNA-Dependent Protein Kinase on the Molecular Fate of the rAAV2 Genome in Skeletal Muscle

• Published in: Proceedings of the National Academy of Sciences - PNAS 2001-03, Vol.98 (7), p.4084-4088
• Main Authors: Song, Sihong, Laipis, Philip J., Berns, Kenneth I., Flotte, Terence R.
• Format: Article
• Language: English
• Subjects: a1-antitrypsin; Adeno-associated virus; Animals; Biological Sciences; Circles; Cytomegalovirus; Deoxyribonucleic acid; Dependovirus - drug effects; Dependovirus - genetics; DNA; DNA probes; DNA-Activated Protein Kinase; DNA-Binding Proteins; DNA-dependent protein kinase; Enzymes; episomes; Genetic Vectors; Genome, Viral; Genomes; Mice; Mice, Inbred C57BL; Mice, SCID; Molecular biology; Muscle, Skeletal - drug effects; Muscle, Skeletal - physiology; Muscle, Skeletal - virology; Muscular system; Plasmids; Protein-Serine-Threonine Kinases - pharmacology; Proteins; Skeletal muscle; Skeletal system; Sprains and strains; transgenes
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.061014598

We report here that the DNA-dependent protein kinase (DNA-PK) affects the molecular fate of the recombinant adeno-associated virus (rAAV) genome in skeletal muscle. rAAV-human α1-antitrypsin (rAAV-hAAT) vectors were delivered by intramuscular injection to either C57BL/6 (DNA-PKcs+) or C57BL/6-SCID [severe combined immunodeficient (SCID), DNA-PKcsPKcs-] mice. In both strains, high levels of transgene expression were sustained for up to 1 year after a single injection. Southern blot analysis showed that rAAV genomes persisted as linear episomes for more than 1 year in SCID mice, whereas only circular episomal forms were observed in the C57BL/6 strain. These results indicate that DNA-PK is involved in the formation of circular rAAV episomes.