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Protein delivery into live cells by incubation with an endosomolytic agent.

ABSTRACT: We report that a tetramethylrhodamine-labeled dimer of the cell-penetrating peptide TAT, dfTAT, penetrates live cells by escaping from endosomes with high efficiency. By mediating endosomal leakage, dfTAT also delivers proteins into cultured cells after a simple co-incubation procedure. We achieved cytosolic delivery in several cell lines and primary cells and observed that only a relatively small amount of material remained trapped inside endosomes. Delivery did not require a binding interaction between dfTAT and a protein, multiple molecules could be delivered simultaneously, and delivery could be repeated. dfTAT-mediated delivery did not noticeably affect cell viability, cell proliferation or gene expression. dfTAT-based intracellular delivery should be useful for cell-based assays, cellular imaging applications and the ex vivo manipulation of cells.

SUBMITTER: Erazo-Oliveras A

PROVIDER: S-EPMC4131206 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature

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Protein delivery into live cells by incubation with an endosomolytic agent.

Erazo-Oliveras Alfredo A Najjar Kristina K Dayani Laila L Wang Ting-Yi TY Johnson Gregory A GA Pellois Jean-Philippe JP

Nature methods 20140615 8

We report that a tetramethylrhodamine-labeled dimer of the cell-penetrating peptide TAT, dfTAT, penetrates live cells by escaping from endosomes with high efficiency. By mediating endosomal leakage, dfTAT also delivers proteins into cultured cells after a simple co-incubation procedure. We achieved cytosolic delivery in several cell lines and primary cells and observed that only a relatively small amount of material remained trapped inside endosomes. Delivery did not require a binding interactio ...[more]

PMID: 24930129

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