Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells
Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells
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DOI：
10.1074/jbc.m010625200
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发表时间：
2001-07-13
影响因子：
4.8
通讯作者：
Nakanishi, M
中科院分区：
文献类型：
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作者：
Eguchi, A;Akuta, T;Nakanishi, M
哺乳动物细胞的质膜是合成递送系统阻止基因转移的严密屏障之一。已使用各种试剂通过在酸性条件下破坏内体膜的稳定性来促进基因转移，但它们的效用是有限的，特别是对于体内基因转移。在本文中，我们报道了人类免疫缺陷病毒1型Tat蛋白（Tat肽）的蛋白转导结构域极大地促进了通过膜去稳定的基因转移，我们构建了重组h噬菌体颗粒，其表面展示Tat肽并携带哺乳动物标记基因作为其基因组的一部分（Tat噬菌体），我们证明，当动物细胞短暂暴露于 Tat 噬菌体时，会诱导噬菌体标记基因的显着表达，而对细胞没有有害影响。相反，展示其他功能肽（例如整联蛋白结合结构域或核定位信号）的重组噬菌体不能诱导可检测的标记基因表达。 Tat 噬菌体诱导的标记基因的表达不受内体促性剂的影响，但会部分受到小凹形成抑制剂的影响。这些数据表明，Tat 肽将成为合成递送载体的有用成分，可独立于经典内吞途径促进基因转移。
The plasma membrane of mammalian cells is one of the tight barriers against gene transfer by synthetic delivery systems. Various agents have been used to facilitate gene transfer by destabilizing the endosomal membrane under acidic conditions, but their utility is limited, especially for gene transfer in vivo, In this article, we report that the protein transduction domain of human immunodeficiency virus type 1 Tat protein (Tat peptide) greatly facilitates gene transfer via membrane destabilization, We constructed recombinant h phage particles displaying Tat peptide on their surfaces and carrying mammalian marker genes as part of their genomes (Tat-phage), We demonstrate that, when animal cells are briefly exposed to Tat-phage, significant expression of phage marker genes is induced with no harmful effects to the cells. In contrast, recombinant phage displaying other functional peptides, such as the integrin-binding domain or a nuclear localization signal, could not induce detectable marker gene expression. The expression of marker genes induced by Tat-phage is not affected by endosomotropic agents but is partially impaired by inhibitors of caveolae formation. These data suggest that Tat peptide will become a useful component of synthetic delivery vehicles that promote gene transfer independently of the classical endocytic pathway.