Extrachromosomal recombinant adeno-associated virus vector genomes are primarily responsible for stable liver transduction in (2001).

Content Provider	CiteSeerX
Author	Nakai, Hiroyuki Yant, Stephen R. Storm, Theresa A. Fuess, Sally Meuse, Leonard Kay, A.
Abstract	Recombinant adeno-associated virus (rAAV) vectors stably transduce hepatocytes in experimental animals. Although the vector genomes are found both as extrachromosomes and as chromosomally integrated forms in hepatocytes, the relative proportion of each has not yet been clearly established. Using an in vivo assay based on the induction of hepatocellular regeneration via a surgical two-thirds partial hepatectomy, we have determined the proportion of integrated and extrachromosomal rAAV genomes in mouse livers and their relative contribution to stable gene expression in vivo. Plasma human coagulation factor IX (hF.IX) levels in mice originating from a chromosomally integrated hF.IX-expressing transposon vector remained unchanged with hepatectomy. This was in sharp contrast to what was observed when a surgical partial hepatectomy was performed in mice 6 weeks to 12 months after portal vein injection of a series of hF.IX-expressing rAAV vectors. At doses of 2.4 � 10 11 to 3.0 � 10 11 vector genomes per mouse (n � 12), hF.IX levels and the average number of stably transduced vector genomes per cell decreased by 92 and 86%, respectively, after hepatectomy. In a separate study, one of three mice injected with a higher dose of rAAV had a higher proportion (67%) of integrated genomes, the significance of which is not known. Nevertheless, in general, these results indicate that, in most cases, no more than �10 % of stably transduced genomes integrated into host chromosomes in vivo. Additionally, the results demonstrate that extrachromosomal, not integrated, genomes are the major form of
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Publisher Date	2001-01-01
Access Restriction	Open
Subject Keyword	Vector Genome Stable Liver Transduction Surgical Partial Hepatectomy Integrated Form Mouse Liver Ix-expressing Transposon Vector Ix Level Surgical Two-thirds Partial Hepatectomy Integrated Genome Integrated Hf Experimental Animal Ix-expressing Raav Vector Relative Contribution Average Number Extrachromosomal Raav Genome Sharp Contrast Recombinant Adeno-associated Virus Major Form Separate Study Host Chromosome Hepatocellular Regeneration Stable Gene Expression Relative Proportion Portal Vein Injection Plasma Human Coagulation Factor Ix
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