Generation of cell-permeable artificial zinc finger protein variants

Designed or artificial zinc finger proteins (ZFPs) are one of the most promising DNA-binding proteins that target genomic sequences of interest in vitro and in vivo. Conjugation of other functional domains such as transcriptional regulatory domains and endonucleases to ZFPs provided powerful molecular tools to modulate endogenous gene expression and genetic information. These ZFP variants have been introduced into cells as DNA-encoding ZFP variants by using plasmids or viral vectors. As an alternative delivery method of ZFP variants, we developed cell-permeable ZFP variants by fusing cell-penetrating peptides to ZFP variants. We will describe how to generate cell-permeable artificial ZFP variants and how to examine the cell permeabilities by immunofluorescent staining.