Author：SIAIS
Date：13 April 2015Research Associate Professor Jia Liu from the Laboratory of ADC Chemistry recently published a paper in collaboration with the Scripps Research Institute. This paper, entitled “Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains”, was published in Journal of Visualized Experiments (JoVE) (doi:10.3791/52814).
Due to their modularity and ability to be reprogrammed to recognize a wide range of DNA sequences, Cys2-His2 zinc-finger DNA-binding domains have emerged as useful tools for targeted genome engineering. Like many other DNA-binding proteins, zinc-fingers also possess the innate ability to cross cell membranes. Dr. Liu, teamed up with Dr. Gaj from The Scripps Research Institute, recently demonstrated that this intrinsic cell-permeability could be leveraged for intracellular protein delivery. Genetic fusion of zinc-finger motifs leads to efficient transport of protein and enzyme cargo into a broad range of mammalian cell types. Unlike other protein transduction technologies, delivery via zinc-finger domains does not inhibit enzyme activity and leads to high levels of cytosolic delivery. Here a detailed step-by-step protocol is presented for the implementation of zinc-finger technology for protein delivery into mammalian cells. Key steps for achieving high levels of intracellular zinc-finger-mediated delivery are highlighted and strategies for maximizing the performance of this system are discussed.
“We have seen efficient delivery of a variety of proteins under cell culture condition using this ZiF platform. Now we are exploring its application in animals and, hopefully, can develop some ZiF product of commercial interest.” Jia Liu said.
