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Prasad, KM, Xu, Y, Yang, Z, Toufektsian, MC, Berr, SS and French, BA. Topoisomerase inhibition accelerates gene expression after adeno-associated virus-mediated gene transfer to the mammalian heart. Mol Ther 15: 764-771

Department of Biomedical Engineering, University of Virginia, Charlottesville, Virginia 22903, USA.
Molecular Therapy (Impact Factor: 6.43). 05/2007; 15(4):764-71. DOI: 10.1038/sj.mt.6300071
Source: PubMed

ABSTRACT Utility of adeno-associated virus 2 (AAV2) vectors for cardiac gene therapy is limited by the prolonged lag phase before maximal gene expression. Topoisomerase inhibition can induce AAV2-mediated gene expression in vivo, but with variable success in different tissues. In this study, we demonstrate that topoisomerase inhibition can accelerate AAV2-mediated gene expression in the mouse heart. We used an AAV2 vector expressing firefly luciferase and monitored expression kinetics using non-invasive bioluminescence imaging. In the group receiving vector alone, cardiac luciferase activity was evident from week 2 onward and increased progressively to reach a steady plateau by 9 weeks postinjection. In the group receiving vector and camptothecine (CPT), luciferase expression was evident from days 2 to 4 onward and increased rapidly to reach a steady plateau by 3-4 weeks postinjection, nearly three times faster than in the absence of CPT (P<0.05). Southern blot analysis of AAV2 genomes in cardiac tissue showed rapid conversion of the AAV2 genome from its single-stranded to double-stranded form in CPT-treated mice. Non-invasive determinations of luciferase expression correlated well with in vitro luciferase assays. Direct injection of the AAV2 vector and long-term luciferase gene expression had no detectable effects on normal cardiac function as assessed by magnetic resonance imaging.

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Available from: Stuart Berr, Aug 12, 2014
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    • "Luciferase expression in live mice was non-invasively assessed by in vivo bioluminescence imaging using previously reported methods (Wu et al., 2002; Prasad et al., 2007). Animals were anesthetized and maintained on 1–1.2% isoflurane in oxygen. "
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