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• W3147668659 abstract "Adeno-associated virus (AAV) has shown great promise as a gene transfer vector. However, the incubation time needed to attain significant levels of gene expression is often too long for some clinical applications. Self-complementary AAV (scAAV) enters the cell as double stranded DNA, eliminating the step of second-strand synthesis, proven to be the rate-limiting step for gene expression of single-stranded AAV (ssAAV). The aim of this study was to compare the efficiency of these two types of AAV vectors in the murine myocardium. Four day old CD-1 mice were injected with either of the two AAV constructs, both expressing GFP and packaged into the AAV1 capsid. The animals were held for 4, 6, 11 or 21 days, after which they were euthanized and their hearts were excised. Serial sections of the myocardial tissue were used for real-time PCR quantification of AAV genome copies and for confocal microscopy. Although we observed similar numbers of AAV genomes at each of the different time points present in both the scAAV and the ssAAV infected hearts, microscopic analysis showed expression of GFP as early as 4 days in animals injected with the scAAV, while little or no expression was observed with the ssAAV constructs until day 11. AAV transduction of murine myocardium is therefore significantly enhanced using scAAV constructs. Results and discussion Adeno-associated virus has become an important tool for gene transfer because of its lack of pathogenicity and its ability to express passenger genes for long periods of time. Although potentially safe as a gene therapy vector, this virus exhibits an extended lag period before transgene expression actually occurs. The reason for this delay in expression is the binding of a cellular protein, FKBP52, to the D-sequence within the inverted terminal repeats (ITRs)[1]. Phosphorylated FKBP52 inhibits viral second strand DNA synthesis, needed for transgene expression, consequently leading to delayed transgene expression [13]. As an avenue for bypassing this phenomenon, McCarty et al. have introduced the use of a double-stranded form of AAV[4]. The typical single-stranded AAV (ssAAV) genome is flanked by two, 145 bp ITRs. The 3' ITR serves as the repPublished: 10 December 2007 Genetic Vaccines and Therapy 2007, 5:13 doi:10.1186/1479-0556-5-13 Received: 1 October 2007 Accepted: 10 December 2007 This article is available from: http://www.gvt-journal.com/content/5/1/13 © 2007 Andino et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited." @default.
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• W3147668659 date "2015-01-01" @default.
• W3147668659 modified "2023-09-27" @default.
• W3147668659 title "Genetic Vaccines and Therapy" @default.
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