Design of carrier peptide-oligonucleotide conjugates with rapid membrane translocation and nuclear localization properties
Design of carrier peptide-oligonucleotide conjugates with rapid membrane translocation and nuclear localization properties
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DOI：
10.1006/bbrc.1997.8050
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发表时间：
1998-02-13
影响因子：
3.1
通讯作者：
Heitz, F
中科院分区：
文献类型：
--
作者：
Chaloin, L;Vidal, P;Heitz, F
在固相合成中，含有与核定位信号相关的疏水基序的多肽被不同的连接物分开。疏水序列对应于信号肽序列或GP41融合肽的片段，而亲水序列是核定位信号的序列。这些多肽的C末端带有半胱胺基团，该基团与荧光探针相连。这使得探针的细胞定位可以作为肽序列的函数来确定。然后将标记的多肽与成纤维细胞孵育。使用N-生物素化的衍生物，我们通过间接免疫荧光证实了观察到的定位与多肽的定位相对应。连接子的存在似乎在细胞定位中发挥了作用。其中一种多肽被成功地用于靶向荧光寡核苷酸进入活细胞，显示了改进的多肽-寡脱氧核苷酸偶联物的细胞递送。(C)1998年学术出版社。
Peptides containing a hydrophobic motif associated with a nuclear localization signal separated by various linkers were synthesized in solid phase. The hydrophobic sequence corresponds either to a signal peptide sequence or to a fragment of the fusion peptide of GP41 while the hydrophilic sequence is that of a nuclear localization signal. The C-termini of these peptides bear a cysteamide group that was linked to a fluorescent probe. This allowed the cellular localization of the probe to be determined as a function of the peptide sequences. The labeled peptides were then incubated with fibroblasts. Using N-biotinylated derivatives we confirmed by indirect immunofluorescence that the observed localizations corresponds to those of the peptides. The presence of a linker appears to play a role in the cellular localization. One of these peptides was successfully used to target fluorescent oligodeoxynucleotides into living cells demonstrating improved cell delivery of peptide-oligodeoxynucleotide conjugates. (C) 1998 Academic Press.