Effect of DNA-dependent protein kinase on the molecular fate of the rAAV2 genome in skeletal muscle.
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 alpha1-antitrypsin (rAAV-hAAT) vectors were delivered by intramuscular injection to either C57BL/6 (DNA-PKcs(+)) or C57BL/6-SCID [severe combined immunodeficient (SCID), DNA-PKcs(-)] 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.
Genetics Institute, Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610, USA.
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S
Song
P J
PJ
Laipis
K I
KI
Berns
T R
TR
Flotte
D000818:Animals; D051747:DNA-Activated Protein Kinase; D004268:DNA-Binding Proteins; D000229:Dependovirus / Q000187:drug effects* / Q000235:genetics; D005822:Genetic Vectors; D016679:Genome, Viral; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000187:drug effects / Q000502:physiology / Q000821:virology*; D017346:Protein Serine-Threonine Kinases / Q000494:pharmacology*
D004268:DNA-Binding Proteins; D051747:DNA-Activated Protein Kinase; D017346:Protein Serine-Threonine Kinases