May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Characterization of the Effects of 2'–MOE Antisense Oligonucleotides on mRNA Levels in Mice After Intravitreal Injection
Author Affiliations & Notes
  • S.P. Henry
    Toxicology,
    Isis Pharmaceuticals Inc, Carlsbad, CA
  • Y. Saisin
    Ophthalmology, The Johns Hopkins University School of Medicine, Baltimore, MD
  • H. Akiyama
    Ophthalmology, The Johns Hopkins University School of Medicine, Baltimore, MD
  • E.G. Marcusson
    Pharmacology,
    Isis Pharmaceuticals Inc, Carlsbad, CA
  • T.M. Vincent
    Pharmacology,
    Isis Pharmaceuticals Inc, Carlsbad, CA
  • P.A. Campochiaro
    Ophthalmology, The Johns Hopkins University School of Medicine, Baltimore, MD
  • Footnotes
    Commercial Relationships  S.P. Henry, Isis Pharmaceuticals E; Y. Saisin, None; H. Akiyama, None; E.G. Marcusson, Isis Pharmaceuticals E; T.M. Vincent, Isis Pharmaceuticals E; P.A. Campochiaro, Isis Pharmaceuticals C.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 4659. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      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.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Abstract: : 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.

Keywords: pharmacology • retina • signal transduction: pharmacology/physiology 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×