DfTat

DfTat functions independently of direct binding interactions with cargo; adding negatively charged proteins such as BSA to the in vitro system reduces the efficiency of dfTAT-mediated cytosolic delivery^[2].
DfTat (1-20 µM; 1 h) penetrates the cytoplasm and nucleus of HeLa, Neuro-2a, HDF, and primary porcine intestinal epithelial cells in a concentration-dependent manner^[1].
DfTat (5 µM; 1 h) delivers EGFP into the cytoplasm and nucleus of HeLa, Neuro-2a, and HDF cells; enhances TAT-Cre-mediated recombination in HeLa reporter cells, producing 47% EGFP-positive cells; delivers functional anti-ATP5A antibodies into HeLa cells; and shows minimal cytotoxicity toward HeLa, MCH58, HDF, and Neuro-2a cells, causing only transient transcriptional changes in HDF cells^[1].
DfTat (3 µM) enhances the transcriptional activity of HOXB4 and TAT-HOXB4 in NIH 3T3 reporter cells^[1].
DfTat induces cytoplasmic penetration in cultured cells at concentrations of 5 µM and above, the percentage of positive cells increases over time, and no plasma membrane permeabilization occurs at 20 µM^[2].
DfTat traffics through early and late endosomes and requires the late endosomal compartment for cytosolic penetration^[2].
DfTat can specifically interact with BMP; DfTat can bind to anionic liposomes and then selectively disrupt BMP-containing late endosome-like liposomes, inducing membrane leakage and lipid mixing; an anti-BMP monoclonal antibody can inhibit this liposomal membrane perturbation effect but does not block the binding behavior of DfTat; in cultured cells, this antibody can completely block DfTat-mediated cytosolic penetration and cargo delivery, yet does not affect the enrichment of DfTat within late endosomes^[2].
DfTat mediates nuclear/nucleolar delivery of TMR-k5 cargo in MDA-MB-231 cells^[3].

DfTat (5 µM) requires macropinocytosis and endosomal acidification, and mediates efficient endosomal leakage in HeLa cells; delivery can be repeated in HeLa cells^[1].
DfTat delivers enzymes, peptides, small molecules, and antibody fragments into cells in vitro through endocytosis and endosomal escape, with 5 μM used for various cargoes^[5].