Gaj Thomas, Liu Jia, Anderson Kimberly E, Sirk Shannon J, Barbas Carlos F
The Skaggs Institute for Chemical Biology and the Departments of Chemistry and Cell and Molecular Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
ACS Chem Biol. 2014 Aug 15;9(8):1662-7. doi: 10.1021/cb500282g. Epub 2014 Jun 19.
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.
开发将蛋白质导入细胞的新方法是推进基础研究和治疗应用的核心挑战。我们之前报道过，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内在的细胞通透性。在此，我们证明基因融合的锌指基序可以将蛋白质和酶转运到多种原代和转化的哺乳动物细胞类型中。我们表明，锌指结构域介导蛋白质摄取的效率超过传统的蛋白质转导系统，并且在不影响酶活性的情况下做到这一点。此外，我们证明锌指蛋白主要通过巨胞饮作用进入细胞，并促进高水平的胞质递送。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，也是蛋白质递送的有效试剂。
