Cellular Delivery and Biological Activity of Antisense Oligonucleotides Conjugated to a Targeted Protein Carrier

Targeted delivery can potentially improve the pharmacological effects of antisense and siRNA oligonucleotides. Here, we describe a novel bioconjugation approach to the delivery of splice-shifting antisense oligonucleotides (SSOs). The SSOs are linked to albumin via reversible S−S bonds. The albumin...

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Bibliographic Details
Published in:Bioconjugate chemistry 2008-11, Vol.19 (11), p.2182-2188
Main Authors: Kang, Hyunmin, Alam, Md. Rowshon, Dixit, Vidula, Fisher, Michael, Juliano, Rudy L
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biochemistry
Cell Line, Tumor
Cell Nucleus - drug effects
Cell Nucleus - metabolism
Cellular biology
Endocytosis
Gene expression
Humans
Luciferases - genetics
Luciferases - metabolism
Oligopeptides - metabolism
Oligoribonucleotides, Antisense - genetics
Oligoribonucleotides, Antisense - metabolism
Oligoribonucleotides, Antisense - pharmacology
Oligoribonucleotides, Antisense - toxicity
Polyethylene Glycols - chemistry
Proteins
Sensitivity and Specificity
Serum Albumin - chemistry
Serum Albumin - metabolism
Toxicity
Tumors
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Summary:Targeted delivery can potentially improve the pharmacological effects of antisense and siRNA oligonucleotides. Here, we describe a novel bioconjugation approach to the delivery of splice-shifting antisense oligonucleotides (SSOs). The SSOs are linked to albumin via reversible S−S bonds. The albumin is also conjugated with poly(ethylene glycol) (PEG) chains that terminate in an RGD ligand that selectively binds the αvβ3 integrin. As a test system, we utilized human melanoma cells that express the αvβ3 integrin and that also contain a luciferase reporter gene that can be induced by delivery of SSOs to the cell nucleus. The RGD-PEG-SSO-albumin conjugates were endocytosed by the cells in an RGD-dependent manner; using confocal fluorescence microscopy, evidence was obtained that the SSOs accumulate in the nucleus. The conjugates were able to robustly induce luciferase expression at concentrations in the 25−200 nM range. At these levels, little short-term or long-term toxicity was observed. Thus, the RGD-PEG-albumin conjugates may provide an effective tool for targeted delivery of oligonucleotides to certain cells and tissues.
ISSN:1043-1802
1520-4812
DOI:10.1021/bc800270w