A Novel Cell-penetrating Peptide, M918, for Efficient Delivery of Proteins and Peptide Nucleic Acids

Department of Neurochemistry, Stockholm University, Stockholm, Sweden.
Molecular Therapy (Impact Factor: 6.23). 11/2007; 15(10):1820-6. DOI: 10.1038/
Source: PubMed


Cell-penetrating peptides (CPPs) have attracted increasing attention in the past decade as a result of their high potential to convey various, otherwise impermeable, bioactive agents across cellular plasma membranes. Albeit different CPPs have proven potent in delivery of different cargoes, there is generally a correlation between high efficacy and cytotoxicity for these peptides. Hence, it is of great importance to find new, non-toxic CPPs with more widespread delivery properties. We present a novel CPP, M918, that efficiently translocates various cells in a non-toxic fashion. In line with most other CPPs, the peptide is internalized mainly via endocytosis, and in particular macropinocytosis, but independent of glycosaminoglycans on the cell surface. In addition, in a splice correction assay using antisense peptide nucleic acid (PNA) conjugated via a disulphide bridge to M918 (M918-PNA), we observed a dose-dependent increase in correct splicing, exceeding the effect of other CPPs. Our data demonstrate that M918 is a novel CPP that can be used to translocate different cargoes inside various cells efficiently.

Download full-text


Available from: Henrik J Johansson, Feb 26, 2015
  • Source
    • "Cell penetrating peptide Luciferase pre-mRNA splicing modulation [30] "
    [Show abstract] [Hide abstract]
    ABSTRACT: Peptides are versatile and attractive biomolecules that can be applied to modulate genetic mechanisms like alternative splicing. In this process, a single transcript yields different mature RNAs leading to the production of protein isoforms with diverse or even antagonistic functions. During splicing events, errors can be caused either by mutations present in the genome or by defects or imbalances in regulatory protein factors. In any case, defects in alternative splicing have been related to several genetic diseases including muscular dystrophy, Alzheimer's disease and cancer from almost every origin. One of the most effective approaches to redirect alternative splicing events has been to attach cell-penetrating peptides to oligonucleotides that can modulate a single splicing event and restore correct gene expression. Here, we summarize how natural existing and bioengineered peptides have been applied over the last few years to regulate alternative splicing and genetic expression. Under different genetic and cellular backgrounds, peptides have been shown to function as potent vehicles for splice correction, and their therapeutic benefits have reached clinical trials and patenting stages, emphasizing the use of regulatory peptides as an exciting therapeutic tool for the treatment of different genetic diseases. Copyright © 2015. Published by Elsevier Inc.
    Full-text · Article · Mar 2015 · Peptides
  • Source
    • "MOEs are RNA analogues formed by modifying the 2′ position in the ribose sugar with the methoxyethyl group and by replacing the phosphodiester bond of the ribose backbone with a phosphorothioate bond, which is stable and resistant to nucleases and imparts high binding affinity and sequence specificity [21]. MOEs have been successfully used to down-regulate various targeted mRNAs via an RNase H-dependant pathway in the form of MOE-DNA gapmers [22], [23], [24], [25], [26], [27], and furthermore it has shown potential in mediating splicing in other models [28], [29], whereas their potential in mediating splice correction in DMD remains to be exploited. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Antisense oligonucleotide (AO)-mediated exon-skipping therapy is one of the most promising therapeutic strategies for Duchenne Muscular Dystrophy (DMD) and several AO chemistries have been rigorously investigated. In this report, we focused on the effect of 2'-O-methoxyethyl oligonucleotides (MOE) on exon skipping in cultured mdx myoblasts and mice. Efficient dose-dependent skipping of targeted exon 23 was achieved in myoblasts with MOE AOs of different lengths and backbone chemistries. Furthermore, we established that 25-mer MOE phosphorothioate (PS) AOs provided the greatest exon-skipping efficacy. When compared with 2'O methyl phosphorothioate (2'OmePS) AOs, 25-mer MOE (PS) AOs also showed higher exon-skipping activity in vitro and in mdx mice after intramuscular injections. Characterization of uptake in vitro corroborated with exon-skipping results, suggesting that increased uptake of 25-mer MOE PS AOs might partly contribute to the difference in exon-skipping activity observed in vitro and in mdx mice. Our findings demonstrate the substantial potential for MOE PS AOs as an alternative option for the treatment of DMD.
    Full-text · Article · Apr 2013 · PLoS ONE
  • Source
    • "The same uptake mechanisms were observed in the absence or presence of cargo molecules. The splice correction assay confirms translocation and bioavailability of the cargo attached to the peptide [14]. Penetratin, the fragment of Antennapedia homeodomain with 16 residues is one of the most extensively used and studied CPPs [7]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Recently, much attention has been given to the problem of drug delivery through the cell-membrane in order to treat and manage several diseases. The discovery of cell penetrating peptides (CPPs) represents a major breakthrough for the transport of large-cargo molecules that may be useful in clinical applications. CPPs are rich in basic amino acids such as arginine and lysine and are able to translocate over membranes and gain access to the cell interior. They can deliver large-cargo molecules, such as oligonucleotides, into cells. Endocytosis and direct penetration have been suggested as the two major uptake mechanisms, a subject still under debate. Unresolved questions include the detailed molecular uptake mechanism(s), reasons for cell toxicity, and the delivery efficiency of CPPs for different cargoes. Here, we give a review focused on uptake mechanisms used by CPPs for membrane translocation and certain experimental factors that affect the mechanism(s).
    Full-text · Article · Apr 2011 · Journal of Biophysics
Show more