Delivery of Proteins, Peptides or Cell-impermeable Small Molecules into Live Cells by Incubation with the Endosomolytic Reagent dfTAT

The overall goal of the following experiment is to deliver macromolecules into live cells using dimer, fluorescent tat or df tat a reagent that can penetrate cells very efficiently without damaging them. This is achieved by first generating and purifying the delivery agent. Dfta as a second step.

Dfta is incubated with cells for one hour at 37 degrees Celsius in the presence of the macromolecule cargo of interest, df tat induces the uptake of the cargo inside Endocytic vesicles. Vesicles mature into early endosomes, finally reaching the late endosome stage where DF tat is able to release the cargo into the cytosolic space of cells. Next, the cells are imaged by fluorescence microscopy in order to assess the delivery efficiency of detta and the cargo of interest.