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

Angew Chem Int Ed Engl. 2015 Dec 7;54(50):15105-8. doi: 10.1002/anie.201505913. Epub 2015 Oct 30.

Abstract

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.

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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell-Penetrating Peptides / analysis*
  • Cell-Penetrating Peptides / metabolism
  • Cytosol / chemistry
  • Cytosol / metabolism*
  • Fluorescence
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / metabolism*
  • HEK293 Cells
  • Humans

Substances

  • Cell-Penetrating Peptides
  • Green Fluorescent Proteins