Adeno-Associated Virus Vectors: Versatile Tools for in vivo Gene Transfer

2008 ◽  
pp. 63-77 ◽  
Author(s):  
Lorena Zentilin ◽  
Mauro Giacca
Keyword(s):  
Gene Transfer ◽  
Adeno Associated Virus ◽  
Virus Vectors ◽  
In Vivo Gene Transfer

scholarly journals Nonrandom Transduction of Recombinant Adeno-Associated Virus Vectors in Mouse Hepatocytes In Vivo: Cell Cycling Does Not Influence Hepatocyte Transduction

2000 ◽  
Vol 74 (8) ◽  
pp. 3793-3803 ◽  
Author(s):  
Carol H. Miao ◽  
Hiroyuki Nakai ◽  
Arthur R. Thompson ◽  
Theresa A. Storm ◽  
Winnie Chiu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Gene Transfer ◽  
Genetic Diseases ◽  
Critical Event ◽  
Dimer Formation ◽  
Adeno Associated Virus ◽  
Virus Vectors ◽  
Preclinical Trials

ABSTRACT Recombinant adeno-associated virus vectors (rAAV) show promise in preclinical trials for the treatment of genetic diseases including hemophilia. Liver-directed gene transfer results in a slow rise in transgene expression, reaching steady-state levels over a period of 5 weeks concomitant with the conversion of the single-stranded rAAV molecules into high-molecular-weight concatemers in about 5% of hepatocytes. Immunohistochemistry and RNA in situ hybridization show that the transgene product is made in about ∼5% of hepatocytes, suggesting that most rAAV-mediated gene expression occurs in hepatocytes containing the double-stranded concatemers. In this study, the mechanism(s) involved in stable transduction in vivo was evaluated. While only ∼5% of hepatocytes are stably transduced, in situ hybridization experiments demonstrated that the vast majority of the hepatocytes take up AAV-DNA genomes after portal vein infusion of the vector. Two different vectors were infused together or staggered by 1, 3, or 5 weeks, and two-color fluorescent in situ hybridization and molecular analyses were performed 5 weeks after the infusion of the second vector. These experiments revealed that a small but changing subpopulation of hepatocytes were permissive to stable transduction. Furthermore, in animals that received a single infusion of two vectors, about one-third of the transduced cells contained heteroconcatemers, suggesting that dimer formation was a critical event in the process of concatemer formation. To determine if the progression through the cell cycle was important for rAAV transduction, animals were continuously infused with 5′-bromo-2′-deoxyuridine (BrdU), starting at the time of administration of a rAAV vector that expressed cytoplasmic β-galactosidase. Colabeling for β-galactosidase and BrdU revealed that there was no preference for transduction of cycling cells. This was further confirmed by demonstrating no increase in rAAV transduction efficiencies in animals whose livers were induced to cycle at the time of or after vector administration. Taken together, our studies suggest that while virtually all hepatocytes take up vector, unknown cellular factors are required for stable transduction, and that dimer formation is a critical event in the transduction pathway. These studies have important implications for understanding the mechanism of integration and may be useful for improving liver gene transfer in vivo.

Successful Gene Transfer Using Adeno-Associated Virus Vectors into the Kidney: Comparison among Adeno-Associated Virus Serotype 1–5 Vectors in vitro and in vivo

2004 ◽  
Vol 96 (4) ◽  
pp. e119-e126 ◽  
Author(s):  
Shin-ichi Takeda ◽  
Masafumi Takahashi ◽  
Hiroaki Mizukami ◽  
Eiji Kobayashi ◽  
Koichi Takeuchi ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Adeno Associated Virus ◽  
Virus Vectors ◽  
Virus Serotype ◽  
Serotype 1

1860: Generation of Fluorescent Sperm by Use of in VIVO Gene Transfer to Hamster Testes

2007 ◽  
Vol 177 (4S) ◽  
pp. 617-618
Author(s):  
Hiraki Kubota ◽  
Kevin Coward ◽  
Olivia Hibbitt ◽  
Nilendran Prathalingam ◽  
William Holt ◽  
...  
Keyword(s):  
Gene Transfer ◽  
In Vivo Gene Transfer
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