title: Protein delivery using Cys2-His2 zinc-finger domains.
abstract: 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.
authors.affiliation: 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.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gaj
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.forename: Kimberly E
authors.initials: KE
authors.lastname: Anderson
authors.forename: Shannon J
authors.initials: SJ
authors.lastname: Sirk
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Barbas
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011506:Proteins / Q000008:administration & dosage* / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: D011506:Proteins; D006639:Histidine; D003545:Cysteine