Combination of a new generation of PNAs with a peptide-based carrier enables efficient targeting of cell cycle progression

Abstract

The design of potent systems for the delivery of charged and noncharged molecules that target genes of interest remains a challenge. We describe a novel technology that combines a new generation of peptide nucleic acids (PNAs), or HypNApPNAs, with a new noncovalent peptide-based delivery system, Pep-2, which promotes efficient delivery of PNAs into several cell lines. We have validated the potential of this technology by showing that Pep2-mediated delivery of an antisense HypNApPNA chimera directed specifically against cyclin B1 induces rapid and robust downregulation of its protein levels and efficiently blocks cell cycle progression of several cell lines, as well as proliferation of cells derived from a breast cancer. Pep-2-based delivery system was shown to be 100-fold more efficient in delivering HypNA-pPNAs than classical cationic lipidbased methods. Whereas Pep-2 is essential for improving the bioavailability of PNAs and HypNA-pPNAs, the latter contribute significantly to the efficiency and specificity of the biological response. We have found that Pep-2/HypNA-pPNA strategy promotes potent antisense effects, which are approximately 25-fold greater than with classical antisense oligonucleotide directed specifically against the same cyclin B1 target. Taken together, these data demonstrate that peptide-mediated delivery of HypNA-pPNAs constitutes a very promising technology for therapeutic applications.

Cell-penetrating peptides: tools for intracellular delivery of therapeutics

“…Another type of primary amphipathic peptide, Pep, was recently developed. It differs from MPG in the nature of the hydrophobic sequence and was designed with the aim of delivering peptides, proteins or PNAs [17,92]. Pep-1, the leader peptide of the Pep family, forms complexes with proteic cargoes which are rapidly delivered into a great variety of cell lines and applied in cellulo and in vivo [93,94].…”

Membrane transducing activity of recombinant Hoxc8 protein and its possible application as a gene delivery vector

“…Not only these we can not exclude the possibility of DNA degradation before and/or after entering the cell. Thus far, there have been many reports on PTD as a delivery vector for proteins though, not many cases have been described for nucleic acid delivery (Morris et al, 2000(Morris et al, , 2004Mukherjee et al, 2005). Although the transfection efficiency was not as effective as commercialized lipofectamine or viral vectors (Harada et al, 2006), using PTDs in combination with other gene delivery systems could enhance the transfection efficiency of PTDs in the future.…”

PNA2/DNA Triplexes: Stability and Specificity

“…To be transported into the cell, PNA is conjugated with a carrier (e.g., peptide) that enters the cell through the cell membrane. Considerable progress has been made in this field by now [35,36]. In addition, PNA was used as a link between a transport peptide and a double-stranded oligonucleotide, which was thereby transferred across the cell membrane [37].…”