Nucleic Acids Research Advance Access originally published online on August 1, 2007
Nucleic Acids Research 2007 35(15):5182-5191; doi:10.1093/nar/gkm478
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Nucleic Acids Research, 2007, Vol. 35, No. 15 5182-5191
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Molecular Biology |
Cell-penetrating peptides as transporters for morpholino oligomers: effects of amino acid composition on intracellular delivery and cytotoxicity
AVI BioPharma, Inc., Corvallis, OR 97333, USA
*To whom correspondence should be addressed. Tel: +1-541-753-3635; Fax: +1-541-754-3545; Email: moulton{at}avibio.com
Received March 13, 2007. Revised May 30, 2007. Accepted May 31, 2007.
Arginine-rich cell-penetrating peptides (CPPs) are promising transporters for intracellular delivery of antisense morpholino oligomers (PMO). Here, we determined the effect of L-arginine, D-arginine and non-
amino acids on cellular uptake, splice-correction activity, cellular toxicity and serum binding for 24 CPP–PMOs. Insertion of 6-aminohexanoic acid (X) or ß-alanine (B) residues into oligoarginine R8 decreased the cellular uptake but increased the splice-correction activity of the resulting compound, with a greater increase for the sequences containing more X residues. Cellular toxicity was not observed for any of the conjugates up to 10 µM. Up to 60 µM, only the conjugates with 
5 Xs exhibited time- and concentration-dependent toxicity. Substitution of L-arginine with D-arginine did not increase uptake or splice-correction activity. High concentration of serum significantly decreased the uptake and splice-correction activity of oligoarginine conjugates, but had much less effect on the conjugates containing X or B. In summary, incorporation of X/B into oligoarginine enhanced the antisense activity and serum-binding profile of CPP–PMO. Toxicity of X/B-containing conjugates was affected by the number of Xs, treatment time and concentration. More active, stable and less toxic CPPs can be designed by optimizing the position and number of R, D-R, X and B residues.
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