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Short antisense oligonucleotide-mediated inhibition is strongly dependent on oligo length and concentration but almost independent of location of the target sequence

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Fakler,  Bernd
Max Planck Institute for Medical Research, Max Planck Society;

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Herlitze,  Stefan
Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society;

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Ruppersberg,  J. Peter
Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society;

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Fakler, B., Herlitze, S., Amthor, B., Zenner, H., & Ruppersberg, J. P. (1994). Short antisense oligonucleotide-mediated inhibition is strongly dependent on oligo length and concentration but almost independent of location of the target sequence. The Journal of Biological Chemistry, 269(23), 16187-16194. Retrieved from http://www.jbc.org/cgi/content/abstract/269/23/16187?maxtoshow%3D%26HITS%3D10%26hits%3D10%26RESULTFORMAT%3D1%26author1%3Druppersberg%252C%2Bj%26searchid%3D1054379177270_599%26stored_search%3D%26FIRSTINDEX%3D0%26flag%3D%26sortspec%3Drelevance%26volume%3D269%26firstpage%3D16187.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0019-A908-3
Abstract
The inhibitory effect of short antisense oligodeoxynucleotides (aODNs) on cRNA expression in Xenopus oocytes was measured using an electrophysiological assay based on subunit-specific block of cloned alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate receptors. The effect of both phosphorothioate-modified (PS) and phosphodiester (PO) aODNs was strongly length dependent with a half-maximal inhibition calculated for an oligo length of 7.6 nucleotides (nt) and 9.9 nt, respectively. More than 95% inhibition was mediated by a PS aODN of 12 nt and by PO aODNs > or = 15 nt. At a given length PS and PO aODNs showed differential dependence of their inhibitory effect on the injected aODN concentration (half-maximal inhibition at 18 ng/microliter for a PO 12-mer and at 0.19 ng/microliter for a PS 12-mer) and differential saturation behavior. The inhibitory effect of aODNs, even as short as 8 nt for PS oligomers, was highly sequence specific, but almost independent of the position of the respective target site on the cRNA (for PS 8-mers, > or = 70% expression inhibition throughout the tested target sites from the translation initiation to the 3'-untranslated region). Thus, short PS aODNs can be reliably used in order to specifically inhibit protein expression in experiments addressing physiological, molecular biological, and perhaps even therapeutical issues.