Detecting Cytosolic Peptide Delivery with the GFP Complementation Assay in the Low Micromolar Range
Schmidt, S., Adjobo-Hermans, M.J.W., Wallbrecher, R., Verdurmen, W.P. R., Bovée-Geurts, P.H.M., van Oostrum, J., Milletti, F., Enderle, T., Brock, R.
Transfection of cells with a plasmid encoding for the first ten strands of the GFP protein (GFP1-10) provides the means to detect cytosolic peptide import at low micromolar concentrations. Cytosolic import of the eleventh strand of the GFP protein either by electroporation or by cell-penetrating peptide-mediated import leads to formation of the full-length GFP protein and fluorescence. An increase in sensitivity is achieved through structural modifications of the peptide and the expression of GFP1-10 as a fusion protein with mCherry.