Journal
JOURNAL OF CONTROLLED RELEASE
Volume 110, Issue 3, Pages 595-604Publisher
ELSEVIER
DOI: 10.1016/j.jconrel.2005.10.026
Keywords
conjugation; oligolysine; PNA; splicing correction; endocytosis
Funding
- Medical Research Council [MC_U105178803] Funding Source: researchfish
- MRC [MC_U105178803] Funding Source: UKRI
- Medical Research Council [MC_U105178803] Funding Source: Medline
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Splicing correction by steric-blocking oligonucleotides (ON) might lead to important clinical applications but requires efficient delivery to cell nuclei. The conjugation of short oligolysine tails has been used to deliver a correcting peptide nucleic acid (PNA) sequence in a positive readout assay in which ON hybridization to the cryptic splice site is strictly required for the expression of a luciferase reporter gene. We have investigated the mechanism of cellular uptake and the efficiency of a (Lys)(8)-PNA-Lys construction in this model system. Cell uptake is temperature-dependent and leads to sequestration of the conjugate in cytoplasmic vesicles in keeping with an endocytic mechanism of internalization. Accordingly a significant and sequence-specific splicing correction is achieved only in the presence of endosome-disrupting agents as chloroquine or 0.5 M sucrose. These endosome-disrupting agents do not affect the activity of free PNA, and do not increase (Lys)(8)-PNA-Lys uptake. (C) 2005 Elsevier B.V. All rights reserved.
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