A practical approach for intracellular protein delivery
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Protein delivery represents a powerful tool for experiments in live cells including studies of protein-protein interactions, protein interference with blocking antibodies, intracellular trafficking and protein or peptide biological functions. Most available reagents dedicated to the protein delivery allow efficient crossing of the plasma membrane. Nevertheless, the major disadvantage for these reagents is a weak release of the delivered protein into the cytoplasm. In this publication we demonstrate efficient protein delivery with a non-peptide based reagent, in human epithelial carcinoma HeLa cells and primary human skin fibroblasts. Using a fluorescent protein in combination with fluorescence microscopy and fluorescence-assisted cell sorting analysis, we show that the delivered protein is indeed released effectively in the cytoplasm, as expected for a dedicated carrier. Furthermore, we present a step-by-step method to optimize conditions for successful intracellular protein delivery.
KeywordsIntracellular release Live cells Non-viral technology Protein delivery Synthetic delivery reagent
Bovine serum albumin
Dynamic light scattering
Fluorescence-assisted cell sorting
Protein transduction domain
We are grateful to Drs C. Muller and J. Peluso for their help in FACS analysis and to Drs J.-P. Behr and J.-F. Williamson for critical reading of the manuscript.
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