A versatile cell-penetrating peptide-adaptor system for efficient delivery of molecular cargos to subcellular destinations.

Scott J. Nowak,Jonathan L. McMurry,Verra M. Ngwa,Maxim Antopolsky,Carol A. Chrestensen,David S. Axford,Allison N. Healey

doi:10.1371/journal.pone.0178648

Scott J. Nowak, Jonathan L. McMurry + Show 5 more

Open Access

https://doi.org/10.1371/journal.pone.0178648

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Journal: PloS one	Publication Date: May 26, 2017
Citations: 25	License type: CC BY 4.0

Affiliation: Kennesaw State University

Abstract

Cell penetrating peptides have long held great potential for delivery of biomolecular cargos for research, therapeutic and diagnostic purposes. They allow rapid, relatively nontoxic passage of a wide variety of biomolecules through the plasma membranes of living cells. However, CPP-based research tools and therapeutics have been stymied by poor efficiency in release from endosomes and a great deal of effort has been made to solve this ‘endosomal escape problem.’ Previously, we showed that use of a reversible, noncovalent coupling between CPP and cargo using calmodulin and a calmodulin binding motif allowed efficient delivery of cargo proteins to the cytoplasm in baby hamster kidney and other mammalian cell lines. The present report demonstrates the efficacy of our CPP-adaptor scheme for efficient delivery of model cargos to the cytoplasm using a variety of CPPs and adaptors. Effective overcoming of the endosomal escape problem is further demonstrated by the delivery of cargo to the nucleus, endoplasmic reticulum and peroxisomes by addition of appropriate subcellular localization signals to the cargos. CPP-adaptors were also used to deliver cargo to myotubes, demonstrating the feasibility of the system as an alternative to transfection for the manipulation of hard-to-transfect cells.

Highlights

Cell-penetrating peptides (CPPs, known as protein transduction domains, or PTDs) allow transport of biomolecular cargo into an array of eukaryotic cells
Discovered some decades ago, there has been a great deal of interest ever since, including development of an array of CPP moieties [1, 2] and strategies for linkage to cargo
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Summary

Introduction

Cell-penetrating peptides (CPPs, known as protein transduction domains, or PTDs) allow transport of biomolecular cargo into an array of eukaryotic cells. Our prior report [15] was the first to use a specific, high-affinity, reversible noncovalent linkage to mediate cargo attachment to CPP moiety, overcoming the endosomal escape problem in a novel way. By addition of localization signals to cargo proteins, delivery to the nucleus, peroxisomes and endoplasmic reticulum was achieved.

Results

Conclusion