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S.P. Henry, Y. Saisin, H. Akiyama, E.G. Marcusson, T.M. Vincent, P.A. Campochiaro; Characterization of the Effects of 2'–MOE Antisense Oligonucleotides on mRNA Levels in Mice After Intravitreal Injection . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4659.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Investigate the effect of intravitreal injection of 2'–Methoxyethoxy modified antisense oligonucleotides on mRNA expression in mouse eyes Methods: Mice were treated with antisense oligonucleotides by a single intravitreal injection. Expression of mRNA of specific genes was assessed by rtPCR evaluation of whole eye or dissected retina, relative to saline control or unrelated oligonucleotide. Sequence specificity of antisense effects were addressed by administering inhibitors for C–raf kinase, Integrin–beta3, PI–4 5–kinase, and ERK–6. Experiments were performed to investigate the dose–response and time–course. Doses ranged from 0.03 to 14 micro–g using a 1 micro–L injection volume. Time points evaluated ranged from 3 to 28 days after injection. Results: A 14 micro–g dose of each of the inhibitors produced sequence–specific reduction in mRNA levels 7 days after a single injection. Levels of mRNA in treated eyes ranged from 40 to 60% of saline or control oligonucleotide levels. The magnitude of reduction appeared greater in dissected retina compared to whole eye, consistent with the regional distribution of oligonucleotide in eye after intravitreal injection. Analysis of the dose response indicated that doses as low as 0.3 micro–g were capable of decreasing expression. Levels of mRNA were decreased by 3 days after injection and remained decreased up to 28 days after injection. Conclusions: Antisense oligonucleotides decreased target gene expression in a sequence dependent fashion after intravitreal injection. Decreases in expression were dose–dependent and prolonged. The antisense inhibitors examined in these studies are of potential interest in studying the mechanism of or potential therapeutic utility in ocular angiogenesis.
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