Calcium ions effectively enhance the effect of antisense peptide nucleic acids conjugated to cationic tat and oligoarginine peptides
Calcium ions effectively enhance the effect of antisense peptide nucleic acids conjugated to cationic tat and oligoarginine peptides
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DOI：
10.1016/j.chembiol.2005.06.009
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发表时间：
2005-08-01
影响因子：
--
通讯作者：
Nielsen, PE
中科院分区：
文献类型：
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作者：
Shiraishi, T;Pankratova, S;Nielsen, PE
细胞穿透性多肽已被广泛用于改善各种蛋白质和反义剂的细胞转运。然而，最近的研究表明，这种阳离子多肽主要是通过内体途径进入细胞的。我们现在发现，加入6 mM的钙离子(以及溶酶体营养剂氯喹)，各种肽核酸(PNA)肽结合物的核反义效应在HeLa细胞中显著增强。特别是，TAT(48-60)和七精氨酸偶联的PNA的反义活性分别提高了44倍和8.5倍。有证据表明，这种机制涉及到内体释放。本研究结果表明，CALL可作为阳离子多肽偶联PNA低聚物体外细胞递送的有效促进剂，同时也强调了这类多肽的内体逃逸途径的重要性。
Cell-penetrating peptides have been widely used to improve cellular delivery of a variety of proteins and antisense agents. However, recent studies indicate that such cationic peptides are predominantly entering cells via an endosomal pathway. We now show that the nuclear antisense effect in HeLa cells of a variety of peptide nucleic acid (PNA) peptide conjugates is significantly enhanced by addition of 6 mM Ca2+ (as well as by the lysosomotrophic agent chloroquine). In particular, the antisense activities of Tat(48-60) and heptaarginine-conjugated PNAs were increased 44-fold and 8.5-fold, respectively. Evidence is presented that the mechanism involves endosomal release. The present results show that Call can be used as an effective enhancer for in vitro cellular delivery of cationic peptide-conjugated PNA oligomers, and also emphasize the significance of the endosomal escape route for such peptides.