title: Comparison of the efficiency of complexes based on S4(13)-PV cell-penetrating peptides in plasmid DNA and siRNA delivery.
abstract: The successful application of gene therapy approaches is highly dependent on the efficient delivery of nucleic acids into target cells. In the present study, new peptide-based nonviral systems were developed to enhance plasmid DNA and siRNA delivery, aiming at generating appropriate gene delivery and gene silencing tools for preclinical and clinical application. For this purpose, a new cell-penetrating peptide derived from the wild-type S4(13)-PV peptide was synthesized through the addition of a five-histidine tail to its N-terminus (H5-S4(13)-PV), and its ability to mediate gene expression and gene silencing was evaluated and compared to that of the wild-type peptide. The histidine-enriched peptide, H5-S4(13)-PV, proved to be generally more efficient and less toxic than the wild-type peptide in the delivery of plasmid DNA. In addition, complexes of H5-S4(13)-PV with siRNAs, but not of S4(13)-PV, were efficiently internalized by cells and presented high knockdown activity (63%). Interestingly, systems containing the S4(13)-PV or the H5-S4(13)-PV peptide exhibited superior biological activity when compared to those containing the reverse NLS or scrambled peptides, suggesting that both the cell-penetrating sequence and the NLS of the S4(13)-PV peptide influence the competence of binary and ternary complexes to accomplish nucleic acid delivery. In order to unravel the cancer therapeutic potential of formulations with the histidine-enriched peptide, their efficiency to mediate silencing of the oncogenic protein survivin was evaluated. As opposed to complexes with the wild-type peptide, H5-S4(13)-PV complexes showed the ability to promote a high survivin knockdown at the level of both protein (44%) and mRNA (73%), in HT1080 cells.
authors.affiliation: CNC - Centre for Neuroscience and Cell Biology, University of Coimbra , Portugal.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Cardoso
authors.forename: Sara
authors.initials: S
authors.lastname: Trabulo
authors.forename: Ana L
authors.initials: AL
authors.lastname: Cardoso
authors.forename: Sílvia
authors.initials: S
authors.lastname: Maia
authors.forename: Paula
authors.initials: P
authors.lastname: Gomes
authors.forename: Amália S
authors.initials: AS
authors.lastname: Jurado
authors.forename: Maria C
authors.initials: MC
authors.lastname: Pedroso de Lima
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics; D008081:Liposomes / Q000737:chemistry; D000077022:Survivin; D014162:Transfection / Q000379:methods
chemical_list: C107489:BIRC5 protein, human; D057846:Cell-Penetrating Peptides; D051034:Inhibitor of Apoptosis Proteins; D008081:Liposomes; D000077022:Survivin