4.8 Article

Detecting Cytosolic Peptide Delivery with the GFP Complementation Assay in the Low Micromolar Range

Journal

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 54, Issue 50, Pages 15105-15108

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201505913

Keywords

cell uptake; cell-penetrating peptides; drug delivery; fragment complementation assays; lipid membranes

Funding

  1. Roche postdoctoral program

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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.

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