Endosome trapping limits the efficiency of splicing correction by PNA-oligolysine conjugates

被引：116

作者：

Abes, S

Williams, D

Prevot, P

Thierry, A

Gait, MJ

Lebleu, B

机构：

[1] Univ Montpellier 2, CNRS, UMR 5124, F-34095 Montpellier, France

[2] MRC, Mol Biol Lab, Cambridge CB2 2QH, England

来源：

JOURNAL OF CONTROLLED RELEASE | 2006年 / 110卷 / 03期

基金：

英国医学研究理事会;

关键词：

conjugation; oligolysine; PNA; splicing correction; endocytosis;

D O I：

10.1016/j.jconrel.2005.10.026

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

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.

引用

页码：595 / 604

页数：10

共 27 条

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