Protein delivery using Cys2-His2 zinc-finger domains.
Protein delivery using Cys2-His2 zinc-finger domains.
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DOI：
10.1021/cb500282g
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发表时间：
2014-08-15
影响因子：
4
通讯作者：
Barbas CF 3rd
中科院分区：
文献类型：
--
作者：
Gaj T;Liu J;Anderson KE;Sirk SJ;Barbas CF 3rd
将蛋白质输送到细胞中的新方法的开发是推进基础研究和治疗应用的核心挑战。我们先前报道，由于Cys2-His2锌指结构域的细胞穿透活性，锌指核酸酶蛋白具有固有的细胞通透性。在这里，我们证明了基因融合的锌指基序可以将蛋白质和酶运输到多种原代和转化的哺乳动物细胞类型中。我们发现，锌指结构域以超过传统蛋白质转导系统的效率介导蛋白质摄取，并且不会影响酶的活性。此外，我们还证明了锌指蛋白主要通过巨噬细胞吞噬进入细胞，促进高水平的胞浆递送。这些发现确立了锌指蛋白不仅是靶向基因组工程的有用工具，而且也是蛋白质传递的有效试剂。
The development of new methods for delivering proteins into cells is a central challenge for advancing both basic research and therapeutic applications. We previously reported that zinc-finger nuclease proteins are intrinsically cell-permeable due to the cell-penetrating activity of the Cys2-His2 zinc-finger domain. Here we demonstrate that genetically fused zinc-finger motifs can transport proteins and enzymes into a wide range of primary and transformed mammalian cell types. We show that zinc-finger domains mediate protein uptake at efficiencies that exceed conventional protein transduction systems and do so without compromising enzyme activity. In addition, we demonstrate that zinc-finger proteins enter cells primarily through macropinocytosis and facilitate high levels of cytosolic delivery. These findings establish zinc-finger proteins as not only useful tools for targeted genome engineering but also effective reagents for protein delivery.